Nucleotide sequences which code for the rpoB gene

ABSTRACT

The present invention relates to polynucleotides corresponding to the rpoB gene and which encode the β-subunit of RNA polymerase B, methods of producing L-amino acids, and methods of screening for polynucleotides which encode proteins having activity of the β-subunit of RNA polymerase B.

CROSS-REFERENCE TO RELATED APPLICATION

[0001] The present application claims priority to German Application No.DE10107229.5 filed Feb. 16, 2001, the entire contents of which areincorporated herein by reference.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates to polynucleotides corresponding tothe rpoB gene and which encode the β-subunit of RNA polymerase B,methods of producing L-amino acids, and methods of screening forpolynucleotides which encode proteins having activity of the β-subunitof RNA polymerase B.

[0004] 2. Discussion of the Background

[0005] L-amino acids, especially L-lysine, are used in human medicineand in the pharmaceuticals industry, in the foodstuffs industry and,very especially, in the feeding of animals.

[0006] It is known that amino acids are produced by fermentation ofstrains of coryneform bacteria, especially Corynebacterium glutamicum.Because of their great importance, attempts are continuously being madeto improve the production processes. Improvements to the processes mayconcern measures relating to the fermentation, such as, for example,stirring and oxygen supply, or the composition of the nutrient media,such as, for example, the sugar concentration during the fermentation,or working up to the product form by, for example, ion-exchangechromatography, or the intrinsic performance properties of themicroorganism itself.

[0007] In order to improve the performance properties of suchmicroorganisms, methods of mutagenesis, selection and mutant selectionare employed. Such methods yield strains which are resistant toantimetabolites or are auxotrophic for metabolites that are important interms of regulation, and which produce amino acids.

[0008] For a number of years, methods of recombinant DNA technology havealso been used for improving the strain of L-amino acid-producingstrains of Corynebacterium, by amplifying individual amino acidbiosynthesis genes and studying the effect on amino acid production.

[0009] However, there remains a critical need for improved methods ofproducing L-amino acids and thus for the provision of strains ofbacteria producing higher amounts of L-amino acids. On a commercial orindustrial scale even small improvements in the yield of L-amino acids,or the efficiency of their production, are economically significant.Prior to the present invention, it was not recognized that enhancementof the rpoB gene encoding the β-subunit of RNA polymerase B wouldimprove L-amino acid yields.

SUMMARY OF THE INVENTION

[0010] One object of the present invention, is providing a new processadjuvant for improving the fermentative production of L-amino acids,particularly L-lysine and L-glutamate. Such process adjuvants includeenhanced bacteria, preferably enhanced Coryneform bacteria which expressenhanced levels of the β-subunit of RNA polymerase B which is encoded bythe rpoB gene.

[0011] Thus, another object of the present invention is providing such abacterium, which expresses enhanced amounts of the β-subunit of RNApolymerase B or gene products of the rpoB gene.

[0012] Another object of the present invention is providing a bacterium,preferably a Coryneform bacterium, which expresses a polypeptide thathas an enhanced β-subunit of RNA polymerase B activity.

[0013] Another object of the invention is to provide a nucleotidesequence encoding a polypeptide which has a β-subunit of RNA polymeraseB sequence. One embodiment of such a sequence is the nucleotide sequenceof SEQ ID NO: 1. Other embodiments of such a sequence is the nucleotidesequences of SEQ ID NOS:3 and 5.

[0014] A further object of the invention is a method of making aβ-subunit of RNA polymerase B or an isolated polypeptide having aβ-subunit of RNA polymerase B activity, as well as use of such isolatedpolypeptides in the production of amino acids. One embodiment of such apolypeptide is the polypeptide having the amino acid sequence of SEQ IDNO: 2. Other embodiments of such a sequence is the amino acid sequenceof SEQ ID NOS:4 and 6.

[0015] In one embodiment the invention provides isolated polypeptidescomprising the amino acid sequences in SEQ ID NOS:2, 4 and/or 6.

[0016] Other objects of the invention include methods of detectingnucleic acid sequences homologous to SEQ ID NO: 1, particularly nucleicacid sequences encoding polypeptides that have the activity of aβ-subunit of RNA polymerase B, and methods of making nucleic acidsencoding such polypeptides.

[0017] The above objects highlight certain aspects of the invention.Additional objects, aspects and embodiments of the invention are foundin the following detailed description of the invention.

DETAILED DESCRIPTION OF THE INVENTION

[0018] Unless otherwise defined, all technical and scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art of molecular biology. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described herein. All publications, patent applications,patents, and other references mentioned herein are incorporated byreference in their entirety. In case of conflict, the presentspecification, including definitions, will control. In addition, thematerials, methods, and examples are illustrative only and are notintended to be limiting.

[0019] Reference is made to standard textbooks of molecular biology thatcontain definitions and methods and means for carrying out basictechniques, encompassed by the present invention. See, for example,Maniatis et al., Molecular Cloning: A Laboratory Manual, Cold SpringHarbor Laboratory, New York (1982) and Sambrook et al., MolecularCloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York(1989) and the various references cited therein.

[0020] Where L-amino acids or amino acids are mentioned hereinbelow,they are to be understood as meaning one or more amino acids, includingtheir salts, selected from the group L-asparagine, L-threonine,L-serine, L-glutamate, L-glycine, L-alanine, L-cysteine, L-valine,L-methionine, L-isoleucine, L-leucine, L-tyrosine, L-phenylalanine,L-histidine, L-lysine, L-tryptophan and L-arginine. L-lysine isespecially preferred.

[0021] Where L-lysine or lysine is mentioned hereinbelow, it is to beunderstood as meaning not only the bases but also the salts, such as,for example, lysine monohydrochloride or lysine sulfate.

[0022] The invention provides an isolated polynucleotide from coryneformbacteria, containing a polynucleotide sequence coding for the rpoB gene,selected from the group

[0023] a) polynucleotide that is at least 70% identical with apolynucleotide that codes for a polypeptide containing the amino acidsequence of SEQ ID No. 2,

[0024] b) polynucleotide that codes for a polypeptide containing anamino acid sequence that is at least 70% identical with the amino acidsequence of SEQ ID No. 2,

[0025] c) polynucleotide that is complementary to the polynucleotides ofa) or b), and

[0026] d) polynucleotide containing at least 15 consecutive nucleotidesof the polynucleotide sequence of a), b) or c),

[0027] the polypeptide preferably exhibiting the activity of theβ-subunit of RNA polymerase B.

[0028] The invention also provides the above-mentioned polynucleotide,it preferably being a replicatable DNA containing:

[0029] (i) the nucleotide sequence shown in SEQ ID No. 1, or

[0030] (ii) at least one sequence that corresponds to sequence (i)within the region of the degeneracy of the genetic code, or

[0031] (iii) at least one sequence that hybridizes with the sequencethat is complementary to sequence (i) or (ii), and optionally

[0032] (iv) sense mutations in (i) which are neutral in terms offunction and which do not change the activity of theprotein/polypeptide.

[0033] Finally, the invention also provides polynucleotides selectedfrom the group

[0034] a) polynucleotides containing at least 15 consecutive nucleotidesselected from the nucleotide sequence of SEQ ID No. 1 between positions1 and 701

[0035] b) polynucleotides containing at least 15 consecutive nucleotidesselected from the nucleotide sequence of SEQ ID No. 1 between positions702 and 4199

[0036] c) polynucleotides containing at least 15 consecutive nucleotidesselected from the nucleotide sequence of SEQ ID No. 1 between positions4200 and 5099.

[0037] The invention also provides

[0038] a replicatable polynucleotide, especially DNA, containing thenucleotide sequence as shown in SEQ ID No. 1;

[0039] a polynucleotide that codes for a polypeptide containing theamino acid sequence as shown in SEQ ID No. 2;

[0040] a vector containing the polynucleotide of the invention,especially a shuttle vector or a plasmid vector, and

[0041] coryneform bacteria which contain the vector or in which the rpoBgene has been enhanced.

[0042] The invention also provides polynucleotides consistingsubstantially of a polynucleotide sequence, which are obtainable byscreening, by means of hybridization, a corresponding gene library of acoryneform bacteria that contains the complete gene or parts thereof,using a probe containing the sequence of the polynucleotide of theinvention according to SEQ ID No. 1 or a fragment thereof, and isolatingthe mentioned polynucleotide sequence.

[0043] Polynucleotides that contain the sequences of the invention aresuitable as hybridization probes for RNA, cDNA and DNA, in order toisolate in their complete length nucleic acids or polynucleotides orgenes that code for the β-subunit of RNA polymerase B, or in order toisolate nucleic acids or polynucleotides or genes that are very similarto the sequence of the rpoB gene. They are likewise suitable forincorporation into so-called “arrays”, “micro arrays” or “DNA chips” inorder to detect and determine the corresponding polynucleotides.

[0044] Polynucleotides that contain the sequences of the invention arealso suitable as primers, with the aid of which it is possible, by meansof the polymerase chain reaction (PCR), to produce DNA of genes thatcode for the β-subunit of RNA polymerase B.

[0045] Such oligonucleotides acting as probes or primers contain atleast 25, 26, 27, 28, 29 or 30, preferably at least 20, 21, 22, 23 or24, very especially preferably at least 15, 16, 17, 18 or 19,consecutive nucleotides. Also suitable are oligonucleotides having alength of at least 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40 or of atleast 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50 nucleotides.Oligonucleotides having a length of at least 100, 150, 200, 250 or 300nucleotides may also be suitable.

[0046] “Isolated” means removed from its natural environment.

[0047] “Polynucleotide” generally refers to polyribonucleotides andpolydeoxyribonucleotides, it being possible for the RNA or DNA to beunmodified or modified.

[0048] The polynucleotides of the invention include a polynucleotideaccording to SEQ ID No. 1 or a fragment prepared therefrom, and alsopolynucleotides that are at least especially from 70% to 80%, preferablyat least from 81% to 85%, especially preferably at least from 86% to90%, and very especially preferably at least 91%, 93%, 95%, 97% or 99%,identical with the polynucleotide according to SEQ ID No. 1, or with afragment prepared therefrom.

[0049] “Polypeptides” are to be understood as being peptides or proteinsthat contain two or more amino acids bonded via peptide bonds.

[0050] The polypeptides of the invention include a polypeptide accordingto SEQ ID No. 2, especially those having the biological activity of theβ-subunit of RNA polymerase B, and also those that are at least from 70%to 80%, preferably at least from 81% to 85%, especially preferably atleast from 86% to 90%, and very especially preferably at least 91%, 93%,95%, 97% or 99%, identical with the polypeptide according to SEQ ID No.2 and exhibit the mentioned activity.

[0051] The invention also provides a process for the production, byfermentation, of amino acids selected from the group L-asparagine,L-threonine, L-serine, L-glutamate, L-glycine, L-alanine, L-cysteine,L-valine, L-methionine, L-isoleucine, L-leucine, L-tyrosine,L-phenylalanine, L-histidine, L-lysine, L-tryptophan and L-arginine,using coryneform bacteria which, in particular, already produce aminoacids and in which the nucleotide sequences coding for the rpoB gene areenhanced, especially overexpressed.

[0052] The term “enhancement” in this connection describes theincreasing of the intracellular activity of one or more enzymes orproteins in a microorganism that are coded for by the corresponding DNA,by, for example, increasing the number of copies of the gene or genes,using a strong promoter or using a gene or allele that codes for acorresponding enzyme or protein having a high level of activity, andoptionally by combining those measures.

[0053] The microorganisms provided by the present invention can produceL-amino acids from glucose, saccharose, lactose, fructose, maltose,molasses, starch, cellulose or from glycerol and ethanol. They may berepresentatives of coryneform bacteria, especially of the genusCorynebacterium. In the case of the genus Corynebacterium, specialmention may be made of the species Corynebacterium glutamicum, which isknown to those skilled in the art for its ability to produce L-aminoacids.

[0054] Suitable strains of the genus Corynebacterium, especially of thespecies Corynebacterium glutamicum (C. glutamicum), are especially theknown wild-type strains

[0055]Corynebacterium glutamicum ATCC13032

[0056]Corynebacterium acetoglutamicum ATCC15806

[0057]Corynebacterium acetoacidophilum ATCC13870

[0058]Corynebacterium thermoaminogenes FERM BP-1539

[0059]Corynebacterium melassecola ATCC17965

[0060]Brevibacterium flavum ATCC14067

[0061]Brevibacterium lactofermentum ATCC13869 and

[0062]Brevibacterium divaricatum ATCC14020

[0063] and L-amino acid-producing mutants or strains prepared therefrom,such as, for example, the L-lysine-producing strains

[0064]Corynebacterium glutamicum FERM-P 1709

[0065]Brevibacterium flavum FERM-P 1708

[0066]Brevibacterium lactofermentum FERM-P 1712

[0067]Corynebacterium glutamicum FERM-P 6463

[0068]Corynebacterium glutamicum FERM-P 6464

[0069]Corynebacterium glutamicum DM58-1

[0070]Corynebacterium glutamicum DG52-5

[0071]Corynebacterium glutamicum DSM5714 and

[0072]Corynebacterium glutamicum DSM12866.

[0073] Preferably, a bacterial strain with attenuated expression of arpoB gene that encodes a polypeptide with activity of the βunit of RNApolymerase B will improve amino acid yield at least 1%.

[0074] The inventors have succeeded in isolating the new rpoB gene of C.glutamicum which codes for the β-subunit of RNA polymerase B, which is aβ-subunit of RNA polymerase B.

[0075] In order to isolate the rpoB gene or other genes from C.glutamicum, a gene library of that microorganism in Escherichia coli (E.coli) is first prepared. The preparation of gene libraries is writtendown in generally known textbooks and handbooks. There may be mentionedas an example the textbook of Winnacker: Gene und Klone, Eine Einführungin die Gentechnologie (Verlag Chemie, Weinheim, Germany, 1990) or thehandbook of Sambrook et al.: Molecular Cloning, A Laboratory Manual(Cold Spring Harbor Laboratory Press, 1989). A very well known genelibrary is that of the E. coli K-12 strain W3110, which has beenprepared by Kohara et al. (Cell 50, 495-508 (1987)) in λ-vectors. Batheet al. (Molecular and General Genetics, 252:255-265, 1996) describe agene library of C. glutamicum ATCC13032, which has been prepared withthe aid of the cosmid vector SuperCos I (Wahl et al., 1987, Proceedingsof the National Academy of Sciences USA, 84:2160-2164) in the E. coliK-12 strain NM554 (Raleigh et al., 1988, Nucleic Acids Research16:1563-1575).

[0076] Börmann et al. (Molecular Microbiology 6(3), 317-326) (sic)(1992)) in turn describe a gene library of C. glutamicum ATCC13032 usingthe cosmid pHC79 (Hohn and Collins, Gene 11, 291-298 (1980)).

[0077] For the preparation of a gene library of C. glutamicum in E. coliit is also possible to use plasmids such as pBR322 (Bolivar, LifeSciences, 25, 807-818 (1979)) or pUC9 (Vieira et al., 1982, Gene,19:259-268). Suitable hosts are especially those E. coli strains thatare restriction- and recombination-defective. An example thereof is thestrain DH5αmcr, which has been described by Grant et al. (Proceedings ofthe National Academy of Sciences USA, 87 (1990) 4645-4649). The long DNAfragments cloned with the aid of cosmids can then in turn be subclonedinto customary vectors suitable for sequencing and then sequenced, as isdescribed, for example, in Sanger et al. (Proceedings of the NationalAcademy of Sciences of the United States of America, 74:5463-5467,1977).

[0078] The resulting DNA sequences can then be studied using knownalgorithms or sequence-analysis programs, such as, for example, that ofStaden (Nucleic Acids Research 14, 217-232 (1986)), that of Marck(Nucleic Acids Research 16, 1829-1836 (1988)) or the GCG program ofButler (Methods of Biochemical Analysis 39, 74-97 (1998)).

[0079] The novel DNA sequence of C. glutamicum coding for the rpoB genehas been found and, as SEQ ID No. 1, forms part of the presentinvention. Furthermore, the amino acid sequence of the correspondingprotein has been derived from the present DNA sequence using the methodsdescribed above. The resulting amino acid sequence of the rpoB geneproduct is shown in SEQ ID No. 2. It is known that enzymes belonging tothe host are able to cleave the N-terminal amino acid methionine orformylmethionine of the protein that is formed.

[0080] Coding DNA sequences that result from SEQ ID No. 1 by thedegeneracy of the genetic code also form part of the invention.Likewise, DNA sequences that hybridize with SEQ ID No. 1 or parts of SEQID No. 1 form part of the invention. Furthermore, to those skilled inthe art, conservative amino acid substitutions, such as, for example,the substitution of glycine with alanine or of aspartic acid withglutamic acid, in proteins are known as sense mutations, which do notlead to any fundamental change in the activity of the protein, that isto say are neutral in terms of function. Such mutations are known interalia also as neutral substitutions. It is also known that changes at theN- and/or C-terminus of a protein do not substantially impair itsfunction or may even stabilise it. The person skilled in the art willfind relevant information inter alia in Ben-Bassat et al. (Journal ofBacteriology 169:751-757 (1987)), in O'Regan et al. (Gene 77:237-251(1989)), in Sahin-Toth et al. (Protein Sciences 3:240-247 (1994)), inHochuli et al. (Bio/Technology 6:1321-1325 (1988)) and in knowntextbooks of genetics and molecular biology. Amino acid sequences thatresult in a corresponding manner from SEQ ID No. 2 likewise form part ofthe invention.

[0081] Similarly, DNA sequences that hybridize with SEQ ID No. 1 orparts of SEQ ID No. 1 form part of the invention. Finally, DNA sequencesthat are produced by the polymerase chain reaction (PCR) using primersthat result from SEQ ID No. 1 form part of the invention. Sucholigonucleotides typically have a length of at least 15 nucleotides.

[0082] The person skilled in the art will find instructions on theidentification of DNA sequences by means of hybridization inter alia inthe handbook “The DIG System Users Guide for Filter Hybridization” fromBoehringer Mannheim GmbH (Mannheim, Germany, 1993) and in Liebl et al.(International Journal of Systematic Bacteriology (1991) 41: 255-260).The hybridization takes place under stringent conditions, that is to saythere are formed only hybrids in which the probe and the targetsequence, i.e. the polynucleotides treated with the probe, are at least70% identical. It is known that the stringency of the hybridization,including the washing steps, is influenced or determined by varying thebuffer composition, the temperature and the salt concentration. Thehybridization reaction is preferably carried out with relatively lowstringency as compared with the washing steps (Hybaid HybridisationGuide, Hybaid Limited, Teddington, UK, 1996).

[0083] There may be used for the hybridization reaction, for example, a5×SSC buffer at a temperature of approximately from 50° C. to 68° C. Inthat case, probes may also hybridize with polynucleotides that are lessthan 70% identical with the sequence of the probe. Such hybrids are lessstable and are removed by washing under stringent conditions. That maybe achieved, for example, by lowering the salt concentration to 2×SSCand optionally subsequently to 0.5×SSC (The DIG System User's Guide forFilter Hybridisation, Boehringer Mannheim, Mannheim, Germany, 1995), atemperature of approximately from 500C to 68° C. being set. It isoptionally possible to lower the salt concentration down to 0.1×SSC. Byraising the hybridization temperature stepwise from 50° C. to 68° C. insteps of approximately from 1 to 2° C., it is possible to isolatepolynucleotide fragments that are, for example, at least 70% or at least80% or at least from 90% to 95% identical with the sequence of the probeused. Further instructions for hybridization are commercially availablein the form of so-called kits (e.g. DIG Easy Hyb from Roche DiagnosticsGmbH, Mannheim, Germany, Catalog No. 1603558).

[0084] The person skilled in the art will find instructions on theamplification of DNA sequences with the aid of the polymerase chainreaction (PCR) inter alia in the handbook of Gait: Oligonukleotidesynthesis: A Practical Approach (IRL Press, Oxford, UK, 1984) and inNewton and Graham: PCR (Spektrum Akademischer Verlag, Heidelberg,Germany, 1994).

[0085] It has been found that coryneform bacteria produce amino acids inan improved manner after enhancement of the rpoB gene.

[0086] In order to achieve overexpression, the number of copies of thecorresponding genes can be increased, or the promoter and regulationregion or the ribosome binding site, which is located upstream of thestructural gene, can be mutated. Expression cassettes inserted upstreamof the structural gene have a similar effect. By means of induciblepromoters it is additionally possible to increase the expression in thecourse of the production of amino acids by fermentation. Expression isalso improved by measures to prolong the life of the m-RNA. Furthermore,the enzyme activity is also enhanced by preventing degradation of theenzyme protein. The genes or gene constructs may either be present inplasmids with different numbers of copies or be integrated and amplifiedin the chromosome. Alternatively, overexpression of the genes inquestion may also be achieved by changing the composition of the mediumand the manner in which culturing is carried out.

[0087] The person skilled in the art will find instructions thereon inMartin et al. (Bio/Technology 5, 137-146 (1987)), in Guerrero et al.(Gene 138, 35-41 (1994)), Tsuchiya and Morinaga (Bio/Technology 6,428-430 (1988)), in Eikmanns et al. (Gene 102, 93-98 (1991)), inEuropean patent specification 0 472 869, in U.S. Pat. No. 4,601,893, inSchwarzer and Pühler (Bio/Technology 9, 84-87 (1991), in Reinscheid etal. (Applied and Environmental Microbiology 60, 126-132 (1994)), inLaBarre et al. (Journal of Bacteriology 175, 1001-1007 (1993)), inpatent application WO 96/15246, in Malumbres et al. (Gene 134, 15-24(1993)), in Japanese Offenlegungsschrift JP-A-10-229891, in Jensen andHammer (Biotechnology and Bioengineering 58, 191-195 (1998)), inMakrides (Microbiological Reviews 60:512-538 (1996)) and in knowntextbooks of genetics and molecular biology.

[0088] For the purposes of enhancement, the rpoB gene of the inventionwas overexpressed, for example, with the aid of episomal plasmids.Suitable plasmids are those which are replicated in coryneform bacteria.Many known plasmid vectors, such as, for example, pZ1 (Menkel et al.,Applied and Environmental Microbiology (1989) 64: 549-554), pEKEx1(Eikmanns et al., Gene 102:93-98 (1991)) or pHS2-1 (Sonnen et al., Gene107:69-74 (1991)), are based on the cryptic plasmids pHM1519, pBL1 orpGA1. Other plasmid vectors, such as, for example, those which are basedon pCG4 (U.S. Pat. No. 4,489,160) or pNG2 (Serwold-Davis et al., FEMSMicrobiology Letters 66, 119-124 (1990)) or pAG1 (U.S. Pat. No.5,158,891), may likewise be used.

[0089] Also suitable are those plasmid vectors with the aid of which theprocess of gene amplification by integration into the chromosome can beapplied, as has been described, for example, by Reinscheid et al.(Applied and Environmental Microbiology 60, 126-132 (1994)) for theduplication or amplification of the hom-thrB operon. In that method, thecomplete gene is cloned into a plasmid vector that is able to replicatein a host (typically E. coli), but not in C. glutamicum. Suitablevectors are, for example, pSUP301 (Simon et al., Bio/Technology 1,784-791 (1983)), pK18mob or pK19mob (Schafer et al., Gene 145, 69-73(1994)), PGEM-T (Promega corporation, Madison, Wis., USA), pCR2.1-TOPO(Shuman (1994). Journal of Biological Chemistry 269:32678-32684; U.S.Pat. No. 5,487,993), pCR®Blunt (Invitrogen, Groningen, Netherlands;Bernard et al., Journal of Molecular Biology, 234: 534-541 (1993)), pEM1(Schrumpf et al., 1991, Journal of Bacteriology 173:4510-4516) or pBGS8(Spratt et al., 1986, Gene 41: 337-342). The plasmid vector containingthe gene to be amplified is then transferred to the desired strain of C.glutamicum by conjugation or transformation. The method of conjugationis described, for example, in Schafer et al. (Applied and EnvironmentalMicrobiology 60, 756-759 (1994)). Methods of transformation aredescribed, for example, in Thierbach et al. (Applied Microbiology andBiotechnology 29, 356-362 (1988)), Dunican and Shivnan (Bio/Technology7, 1067-1070 (1989)) and Tauch et al. (FEMS Microbiological Letters 123,343-347 (1994)). After homologous recombination by means of a“cross-over” occurrence, the resulting strain contains at least twocopies of the gene in question.

[0090] It has also been found that the substitution of amino acids,especially in the sections between position 1 to 10, 190 to 200 and 420to 450 in the amino acid sequence of the β-subunit of RNA polymerase Bshown in SEQ ID No. 2, improves the lysine production of coryneformbacteria.

[0091] It has also been found that the substitution of amino acids atone or more positions selected from the group a) position 1 to 10, b)position 190 to 200 and c) position 420 to 450 in SEQ ID No. 2 may takeplace simultaneously.

[0092] In the region between position 1 to 10, preference is given tothe substitution of L-proline at position 5 by L-leucine, L-isoleucineor L-valine.

[0093] In the region between position 190 to 200, preference is given tothe substitution of L-serine at position 196 by L-phenylalanine orL-tyrosine.

[0094] In the region between 420 to 450, the following substitutions arepreferred: substitution of L-leucine at position 424 by L-proline orL-arginine, substitution of L-serine at position 425 by L-threonine orL-alanine, substitution of L-glutamine at position 426 by L-leucine orL-lysine, substitution of L-aspartic acid at position 429 byL-isoleucine, L-valine or L-leucine, substitution of L-histidine atposition 439 by any proteinogenic amino acid with the exception ofL-histidine, is (sic) the substitution of L-serine at position 444 byL-leucine, L-tyrosine or L-tryptophan, and substitution of L-leucine atposition 446 by L-proline or L-isoleucine.

[0095] Very special preference is given to one or more amino acidsubstitutions selected from the group: L-proline at position 5 byL-leucine, L-serine at position 196 by L-phenylalanine, L-aspartate atposition 429 by L-valine, and L-histidine at position 439 by L-tyrosine.

[0096] SEQ ID No. 3 shows the base sequence of the allele rpoB-1547contained in strain DM1547. The rpoB-1547 allele codes for a protein theamino acid sequence of which is shown in SEQ ID No. 4. The proteincontains L-leucine at position 5, L-phenylalanine at position 196 andL-valine at position 429. The DNA sequence of the rpoB-1547 allele (SEQID No. 3) contains the following base substitutions as compared with therpoB wild-type gene (SEQ ID No. 1): thymine at position 715 instead ofcytosine, thymine at position 1288 instead of cytosine, and thymine atposition 1987 instead of adenine.

[0097] SEQ ID No. 5 shows the base sequence of the allele rpoB-1546contained in strain DM1546. The rpoB-1546 allele codes for a protein theamino acid sequence of which is shown in SEQ ID No. 6. The proteincontains L-tyrosine at position 439. The DNA sequence of the rpoB-1546allele (SEQ ID No. 5) contains the following base substitutions ascompared with the rpoB wild-type gene (SEQ ID No. 1): thymine atposition 2016 instead of cytosine.

[0098] There may be employed for the mutagenesis conventional methods ofmutagenesis using mutagenic substances such as, for example,N-methyl-N′-nitro-N-nitrosoguanidine or ultraviolet light. There mayalso be used for the mutagenesis in vitro methods such as, for example,treatment with hydroxylamine (Miller, J. H.: A Short Course in BacterialGenetics. A Laboratory Manual and Handbook for Escherichia coli andRelated Bacteria, Cold Spring Harbor Laboratory Press, Cold SpringHarbor, 1992) or mutagenic oligonucleotides (T. A. Brown: Gentechnologiefür Einsteiger, Spektrum Akademischer Verlag, Heidelberg, 1993) or thepolymerase chain reaction (PCR), as is described in the handbook ofNewton and Graham (PCR, Spektrum Akademischer Verlag, Heidelberg, 1994).

[0099] In addition, it may be advantageous for the production of L-aminoacids to enhance, especially to overexpress, in addition to the rpoBgene, one or more enzymes of the biosynthesis pathway in question, ofglycolysis, of the anaplerotic pathway, of the citric acid cycle, of thepentose phosphate cycle, of amino acid export, and, optionally,regulatory proteins.

[0100] Accordingly, for the production of L-lysine, in addition toenhancing the rpoB gene, one or more genes selected from the group

[0101] the gene dapA coding for dihydrodipicolinate synthase (EP-B 0 197335),

[0102] the gene gap coding for glyceraldehyde 3-phosphate dehydrogenase(Eikmanns (1992), Journal of Bacteriology 174:6076-6086),

[0103] the gene tpi coding for triose phosphate isomerase (Eikmanns(1992), Journal of Bacteriology 174:6076-6086),

[0104] the gene pgk coding for 3-phosphoglycerate kinase (Eikmanns(1992), Journal of Bacteriology 174:6076-6086),

[0105] the gene zwf coding for glucose-6-phosphate dehydrogenase(JP-A-09224661),

[0106] the gene pyc coding for pyruvate carboxylase (DE-A-198 31 609),

[0107] the gene mqo coding for malate quinone oxidoreductase (Molenaaret al., European Journal of Biochemistry 254, 395-403 (1998)),

[0108] the gene lysc coding for a feed-back resistant aspartate kinase(Kalinowski et al., Molecular Microbiologie 5(5), 1197-1204 (1991)),

[0109] the gene lysE coding for lysine export (DE-A-195 48 222),

[0110] the gene zwa1 coding for the Zwa1 protein (DE: 19959328.0, DSM13115), and

[0111] the rpsL gene coding for ribosomal protein S12 and shown in SEQID No. 7 and 8

[0112] may be enhanced, especially overexpressed.

[0113] The term “attenuation” in this connection describes thediminution or exclusion of the intracellular activity of one or moreenzymes (proteins) in a microorganism that are coded for by thecorresponding DNA, by, for example, using a weak promoter or using agene or allele that codes for a corresponding enzyme having lowactivity, or by inactivating the corresponding gene or enzyme (protein),and optionally by combining those measures.

[0114] Furthermore, it may be advantageous for the production of L-aminoacids, in addition to enhancing the rpoB gene, to attenuate, especiallyto diminish the expression of, one or more genes selected from the group

[0115] the gene pck coding for phosphoenol pyruvate carboxykinase (DE199 50 409.1; DSM 13047),

[0116] the gene pgi coding for glucose-6-phosphate isomerase (U.S. Ser.NO. 09/396,478; DSM 12969),

[0117] the gene poxB coding for pyruvate oxidase (DE: 1995 1975.7; DSM13114),

[0118] the gene zwa2 coding for the Zwa2 protein (DE: 19959327.2, DSM13113).

[0119] It may also be advantageous for the production of amino acids, inaddition to enhancing the rpoB gene, to exclude undesired secondaryreactions (Nakayama: “Breeding of Amino Acid Producing Micro-organisms”,in: Overproduction of Microbial Products, Krumphanzl, Sikyta, Vanek(eds.), Academic Press, London, UK, 1982).

[0120] The microorganisms produced according to the invention also formpart of the invention and can be cultivated, for the purposes of theproduction of amino acids, continuously or discontinuously in the batch,fed batch or repeated fed batch process. A summary of known cultivationmethods is described in the textbook of Chmiel (Bioprozeβtechnik 1.Einführung in die Bioverfahrenstechnik (Gustav Fischer Verlag,Stuttgart, 1991)) or in the textbook of Storhas (Bioreaktoren undperiphere Einrichtungen (Vieweg Verlag, Braunschweig/Wiesbaden, 1994)).

[0121] The culture medium to be used must meet the requirements of thestrains in question in a suitable manner. Descriptions of culture mediafor various microorganisms are to be found in the handbook “Manual ofMethods for General Bacteriology” of the American Society forBacteriology (Washington D.C., USA, 1981).

[0122] There may be used as the carbon source sugars and carbohydrates,such as, for example, glucose, saccharose, lactose, fructose, maltose,molasses, starch and cellulose, oils and fats, such as, for example,soybean oil, sunflower oil, groundnut oil and coconut oil, fatty acids,such as, for example, palmitic acid, stearic acid and linoleic acid,alcohols, such as, for example, glycerol and ethanol, and organic acids,such as, for example, acetic acid. Those substances may be usedindividually or in the form of a mixture.

[0123] There may be used as the nitrogen source organicnitrogen-containing compounds, such as peptones, yeast extract, meatextract, malt extract, corn steep liquor, soybean flour and urea, orinorganic compounds, such as ammonium sulfate, ammonium chloride,ammonium phosphate, ammonium carbonate and ammonium nitrate. Thenitrogen sources may be used individually or in the form of a mixture.

[0124] There may be used as the phosphorus source phosphoric acid,potassium dihydrogen phosphate or dipotassium hydrogen phosphate or thecorresponding sodium-containing salts. The culture medium must alsocontain salts of metals, such as, for example, magnesium sulfate or ironsulfate, which are necessary for growth. Finally, essential growthsubstances, such as amino acids and vitamins, may be used in addition tothe above-mentioned substances. Suitable precursors may also be added tothe culture medium. The mentioned substances may be added to the culturein the form of a single batch, or they may be fed in in a suitablemanner during the cultivation.

[0125] In order to control the pH value of the culture, basic compounds,such as sodium hydroxide, potassium hydroxide, ammonia or ammonia water,or acid compounds, such as phosphoric acid or sulfuric acid, areexpediently used. In order to control the development of foam,anti-foams, such as, for example, fatty acid polyglycol esters, may beused. In order to maintain the stability of plasmids, suitablesubstances having a selective action, such as, for example, antibiotics,may be added to the medium. In order to maintain aerobic conditions,oxygen or gas mixtures containing oxygen, such as, for example, air, areintroduced into the culture. The temperature of the culture is normallyfrom 20° C. to 45° C. and preferably from 25° C. to 40° C. The cultureis continued until the maximum amount of the desired product has formed.That aim is normally achieved within a period of from 10 hours to 160hours.

[0126] Methods of determining L-amino acids are known from the priorart. The analysis may be carried out, for example, as described inSpackman et al. (Analytical Chemistry, 30, (1958), 1190) by ion-exchangechromatography with subsequent ninhydrin derivatization, or it may becarried out by reversed phase HPLC, as described in Lindroth et al.(Analytical Chemistry (1979) 51: 1167-1174).

[0127] Pure cultures of the following microorganisms were deposited onJan. 16, 2001 at the Deutsche Sammlung für Mikroorganismen undZellkulturen (DSMZ, Braunschweig, Germany) in accordance with theBudapest Treaty:

[0128]Corynebacterium glutamicum strain DM1546 as DSM 13993

[0129]Corynebacterium glutamicum strain DM1547 as DSM 13994.

[0130] The process of the invention is used for the production of aminoacids by fermentation.

[0131] The present invention is explained in greater detail below bymeans of Examples.

[0132] The isolation of plasmid DNA from Escherichia coli and alltechniques for restriction, Klenow and alkaline phosphatase treatmentwere carried out according to Sambrook et al. (Molecular Cloning. ALaboratory Manual (1989) Cold Spring Harbour Laboratory Press, ColdSpring Harbor, N.Y., USA). Methods for the transformation of Escherichiacoli are also described in that handbook.

[0133] The composition of common nutrient media, such as LB or TYmedium, will also be found in the handbook of Sambrook et al.

EXAMPLE 1

[0134] Preparation of a Genomic Cosmid Gene Library from Corynebacteriumglutamicum ATCC 13032

[0135] Chromosomal DNA from Corynebacterium glutamicum ATCC 13032 isisolated as described in Tauch et al. (1995, Plasmid 33:168-179) andpartially cleaved with the restriction enzyme Sau3AI (AmershamPharmacia, Freiburg, Germany, product description Sau3AI, Code no.27-0913-02). The DNA fragments are dephosphorylated with shrimp alkalinephosphatase (Roche Diagnostics GmbH, Mannheim, Germany, productdescription SAP, Code no. 1758250). The DNA of cosmid vector SuperCosl(Wahl et al. (1987) Proceedings of the National Academy of Sciences USA84:2160-2164), obtained from Stratagene (La Jolla, USA, productdescription SuperCosl Cosmid Vektor Kit, Code no. 251301), is cleavedwith the restriction enzyme XbaI (Amersham Pharmacia, Freiburg, Germany,product description XbaI, Code no. 27-0948-02) and likewisedephosphorylated with shrimp alkaline phosphatase.

[0136] The cosmid DNA is then cleaved with the restriction enzyme BamHI(Amersham Pharmacia, Freiburg, Germany, product description BamHI, Codeno. 27-0868-04). The cosmid DNA so treated is mixed with the treatedATCC13032 DNA, and the batch is treated with T4-DNA ligase (AmershamPharmacia, Freiburg, Germany, product description T4-DNA ligase, Codeno. 27-0870-04). The ligation mixture is then packed in phages with theaid of Gigapack II XL Packing Extract (Stratagene, La Jolla, USA,product description Gigapack II XL Packing Extract, Code no. 200217).

[0137] For infection of E. coli strain NM554 (Raleigh et al. 1988,Nucleic Acid Research 16:1563-1575), the cells are taken up in 10 mMMgSO₄ and mixed with an aliquot of the phage suspension. Infection andtitration of the cosmid library are carried out as described in Sambrooket al. (1989, Molecular Cloning: A Laboratory Manual, Cold SpringHarbor), the cells being plated out on LB agar (Lennox, 1955, Virology,1:190) with 100 mg/l ampicillin. After incubation overnight at 37° C.,recombinant individual clones are selected.

EXAMPLE 2

[0138] Isolation and Sequencing of the rpoB Gene

[0139] The cosmid DNA of an individual colony is isolated using theQiaprep Spin Miniprep Kit (Product No. 27106, Qiagen, Hilden, Germany)according to the manufacturer's instructions, and partially cleaved withthe restriction enzyme Sau3AI (Amersham Pharmacia, Freiburg, Germany,product description Sau3AI, Product No. 27-0913-02). The DNA fragmentsare dephosphorylated with shrimp alkaline phosphatase (Roche DiagnosticsGmbH, Mannheim, Germany, product description SAP, Product No. 1758250).After separation by gel electrophoresis, cosmid fragments having a sizein the range from 1500 to 2000 bp are isolated using the QiaExII GelExtraction Kit (Product No. 20021, Qiagen, Hilden, Germany).

[0140] The DNA of sequencing vector pZero-1, obtained from Invitrogen(Groningen, Netherlands, product description Zero Background CloningKit, Product No. K2500-01), is cleaved with the restriction enzyme BamHI(Amersham Pharmacia, Freiburg, Germany, product description BamHI,Product No. 27-0868-04). Ligation of the cosmid fragments into thesequencing vector pZero-1 is carried out as described by Sambrook et al.(1989, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor), theDNA mixture being incubated overnight with T4 ligase (Pharmacia Biotech,Freiburg, Germany). The ligation mixture is then electroporated into E.coli strain DH5αMCR (Grant, 1990, Proceedings of the National Academy ofSciences U.S.A., 87:4645-4649) (Tauch et al. 1994, FEMS MicrobiolLetters, 123:343-347) and plated out on LB agar (Lennox, 1955, Virology,1:190) with 50 mg/l Zeocin.

[0141] Plasmid preparation of the recombinant clones is carried outusing the Biorobot 9600 (Product No. 900200, Qiagen, Hilden, Germany).Sequencing is effected by the dideoxy chain termination method of Sangeret al. (1977, Proceedings of the National Academy of Sciences U.S.A.,74:5463-5467) with modifications according to Zimmermann et al. (1990,Nucleic Acids Research, 18:1067). The “RR dRhodamin Terminator CycleSequencing Kit” from PE Applied Biosystems (Product No. 403044,Weiterstadt, Germany) is used. Separation by gel electrophoresis andanalysis of the sequencing reaction is carried out in a “Rotiphorese NFAcrylamid/Bisacrylamid” gel (29:1) (Product No. A124.1, Roth, Karlsruhe,Germany) using the “ABI Prism 377” sequencing device from PE AppliedBiosystems (Weiterstadt, Germany).

[0142] The resulting crude sequence data are then processed using theStaden program package (1986, Nucleic Acids Research, 14:217-231)Version 97-0. The individual sequences of the pZero1 derivatives areassembled to a coherent contig. The computer-assisted coding regionanalysis is prepared using the program XNIP (Staden, 1986, Nucleic AcidsResearch, 14:217-231).

[0143] The resulting nucleotide sequence is shown in SEQ ID No. 1.Analysis of the nucleotide sequence gives an open reading frame of 3497base pairs, which is designated the rpoB gene. The rpoB gene codes for aprotein of 1165 amino acids.

[0144] Obviously, numerous modifications and variations on the presentinvention are possible in light of the above teachings. It is thereforeto be understood that within the scope of the appended claims, theinvention may be practiced otherwise than as specifically describedherein.

1 12 1 5099 DNA Corynebacterium glutamicum CDS (702)..(4196) 1acaatgtgac tcgtgatttt tgggtggatc agcgtaccgg tttggttgtc gatctagctg 60aaaatattga tgatttttac ggcgaccgca gcggccagaa gtacgaacag aaattgcttt 120tcgacgcctc cctcgacgat gcagctgtct ctaagctggt tgcacaggcc gaaagcatcc 180ctgatggaga tgtgagcaaa atcgcaaata ccgtaggtat tgtgatcggt gcggtattgg 240ctctcgtggg cctggccggg tgttttgggg cgtttgggaa gaaacgtcga gaagcttaac 300ctgctgttca aatagatttt ccctgtttcg aattgcggaa accccgggtt tgtttgctag 360ggtgcctcgt agaaggggtc aagaagattt ctgggaaacg cgcccgtgcg gttggttgct 420aatagcacgc ggagcaccag atgaaaaatc tcccctttac tttcgcgcgc gattggtata 480ctctgagtcg ttgcgttgga attcgtgact ctttttcgtt cctgtagcgc caagaccttg 540atcaaggtgg tttaaaaaaa ccgatttgac aaggtcattc agtgctatct ggagtcgttc 600agggggatcg ggttcctcag cagaccaatt gctcaaaaat accagcggtg ttgatctgca 660cttaatggcc ttgaccagcc aggtgcaatt acccgcgtga g gtg ctg gaa gga ccc 716Val Leu Glu Gly Pro 1 5 atc ttg gca gtc tcc cgc cag acc aag tca gtc gtcgat att ccc ggt 764 Ile Leu Ala Val Ser Arg Gln Thr Lys Ser Val Val AspIle Pro Gly 10 15 20 gca ccg cag cgt tat tct ttc gcg aag gtg tcc gca cccatt gag gtg 812 Ala Pro Gln Arg Tyr Ser Phe Ala Lys Val Ser Ala Pro IleGlu Val 25 30 35 ccc ggg cta cta gat ctt caa ctg gat tct tac tcc tgg ctgatt ggt 860 Pro Gly Leu Leu Asp Leu Gln Leu Asp Ser Tyr Ser Trp Leu IleGly 40 45 50 acg cct gag tgg cgt gct cgt cag aag gaa gaa ttc ggc gag ggagcc 908 Thr Pro Glu Trp Arg Ala Arg Gln Lys Glu Glu Phe Gly Glu Gly Ala55 60 65 cgc gta acc agc ggc ctt gag aac att ctc gag gag ctc tcc cca atc956 Arg Val Thr Ser Gly Leu Glu Asn Ile Leu Glu Glu Leu Ser Pro Ile 7075 80 85 cag gat tac tct gga aac atg tcc ctg agc ctt tcg gag cca cgc ttc1004 Gln Asp Tyr Ser Gly Asn Met Ser Leu Ser Leu Ser Glu Pro Arg Phe 9095 100 gaa gac gtc aag aac acc att gac gag gcg aaa gaa aag gac atc aac1052 Glu Asp Val Lys Asn Thr Ile Asp Glu Ala Lys Glu Lys Asp Ile Asn 105110 115 tac gcg gcg cca ctg tat gtg acc gcg gag ttc gtc aac aac acc acc1100 Tyr Ala Ala Pro Leu Tyr Val Thr Ala Glu Phe Val Asn Asn Thr Thr 120125 130 ggt gaa atc aag tct cag act gtc ttc atc ggc gat ttc cca atg atg1148 Gly Glu Ile Lys Ser Gln Thr Val Phe Ile Gly Asp Phe Pro Met Met 135140 145 acg gac aag gga acg ttc atc atc aac gga acc gaa cgc gtt gtg gtc1196 Thr Asp Lys Gly Thr Phe Ile Ile Asn Gly Thr Glu Arg Val Val Val 150155 160 165 agc cag ctc gtc cgc tcc ccg ggc gtg tac ttt gac cag acc atcgat 1244 Ser Gln Leu Val Arg Ser Pro Gly Val Tyr Phe Asp Gln Thr Ile Asp170 175 180 aag tca act gag cgt cca ctg cac gcc gtg aag gtt att cct tcccgt 1292 Lys Ser Thr Glu Arg Pro Leu His Ala Val Lys Val Ile Pro Ser Arg185 190 195 ggt gct tgg ctt gag ttt gac gtc gat aag cgc gat tcg gtt ggtgtt 1340 Gly Ala Trp Leu Glu Phe Asp Val Asp Lys Arg Asp Ser Val Gly Val200 205 210 cgt att gac cgc aag cgt cgc cag cca gtc acc gta ctg ctg aaggct 1388 Arg Ile Asp Arg Lys Arg Arg Gln Pro Val Thr Val Leu Leu Lys Ala215 220 225 ctt ggc tgg acc act gag cag atc acc gag cgt ttc ggt ttc tctgaa 1436 Leu Gly Trp Thr Thr Glu Gln Ile Thr Glu Arg Phe Gly Phe Ser Glu230 235 240 245 atc atg atg tcc acc ctc gag tcc gat ggt gta gca aac accgat gag 1484 Ile Met Met Ser Thr Leu Glu Ser Asp Gly Val Ala Asn Thr AspGlu 250 255 260 gca ttg ctg gag atc tac cgc aag cag cgt cca ggc gag cagcct acc 1532 Ala Leu Leu Glu Ile Tyr Arg Lys Gln Arg Pro Gly Glu Gln ProThr 265 270 275 cgc gac ctt gcg cag tcc ctc ctg gac aac agc ttc ttc cgtgca aag 1580 Arg Asp Leu Ala Gln Ser Leu Leu Asp Asn Ser Phe Phe Arg AlaLys 280 285 290 cgc tac gac ctg gct cgc gtt ggt cgt tac aag atc aac cgcaag ctc 1628 Arg Tyr Asp Leu Ala Arg Val Gly Arg Tyr Lys Ile Asn Arg LysLeu 295 300 305 ggc ctt ggt ggc gac cac gat ggt ttg atg act ctt act gaagag gac 1676 Gly Leu Gly Gly Asp His Asp Gly Leu Met Thr Leu Thr Glu GluAsp 310 315 320 325 atc gca acc acc atc gag tac ctg gtg cgt ctg cac gcaggt gag cgc 1724 Ile Ala Thr Thr Ile Glu Tyr Leu Val Arg Leu His Ala GlyGlu Arg 330 335 340 gtc atg act tct cca aat ggt gaa gag atc cca gtc gagacc gat gac 1772 Val Met Thr Ser Pro Asn Gly Glu Glu Ile Pro Val Glu ThrAsp Asp 345 350 355 atc gac cac ttt ggt aac cgt cgt ctg cgt acc gtt ggcgaa ctg atc 1820 Ile Asp His Phe Gly Asn Arg Arg Leu Arg Thr Val Gly GluLeu Ile 360 365 370 cag aac cag gtc cgt gtc ggc ctg tcc cgc atg gag cgcgtt gtt cgt 1868 Gln Asn Gln Val Arg Val Gly Leu Ser Arg Met Glu Arg ValVal Arg 375 380 385 gag cgt atg acc acc cag gat gcg gag tcc att act cctact tcc ttg 1916 Glu Arg Met Thr Thr Gln Asp Ala Glu Ser Ile Thr Pro ThrSer Leu 390 395 400 405 atc aac gtt cgt cct gtc tct gca gct atc cgt gagttc ttc gga act 1964 Ile Asn Val Arg Pro Val Ser Ala Ala Ile Arg Glu PhePhe Gly Thr 410 415 420 tcc cag ctg tct cag ttc atg gac cag aac aac tccctg tct ggt ttg 2012 Ser Gln Leu Ser Gln Phe Met Asp Gln Asn Asn Ser LeuSer Gly Leu 425 430 435 act cac aag cgt cgt ctg tcg gct ctg ggc ccg ggtggt ctg tcc cgt 2060 Thr His Lys Arg Arg Leu Ser Ala Leu Gly Pro Gly GlyLeu Ser Arg 440 445 450 gag cgc gcc ggc atc gag gtt cga gac gtt cac ccatct cac tac ggc 2108 Glu Arg Ala Gly Ile Glu Val Arg Asp Val His Pro SerHis Tyr Gly 455 460 465 cgt atg tgc cca att gag act ccg gaa ggt cca aacatt ggc ctg atc 2156 Arg Met Cys Pro Ile Glu Thr Pro Glu Gly Pro Asn IleGly Leu Ile 470 475 480 485 ggt tcc ttg gct tcc tat gct cga gtg aac ccattc ggt ttc att gag 2204 Gly Ser Leu Ala Ser Tyr Ala Arg Val Asn Pro PheGly Phe Ile Glu 490 495 500 acc cca tac cgt cgc atc atc gac ggc aag ctgacc gac cag att gac 2252 Thr Pro Tyr Arg Arg Ile Ile Asp Gly Lys Leu ThrAsp Gln Ile Asp 505 510 515 tac ctt acc gct gat gag gaa gac cgc ttc gttgtt gcg cag gca aac 2300 Tyr Leu Thr Ala Asp Glu Glu Asp Arg Phe Val ValAla Gln Ala Asn 520 525 530 acg cac tac gac gaa gag ggc aac atc acc gatgag acc gtc act gtt 2348 Thr His Tyr Asp Glu Glu Gly Asn Ile Thr Asp GluThr Val Thr Val 535 540 545 cgt ctg aag gac ggc gac atc gcc atg gtt ggccgc aac gcg gtt gat 2396 Arg Leu Lys Asp Gly Asp Ile Ala Met Val Gly ArgAsn Ala Val Asp 550 555 560 565 tac atg gac gtt tcc cct cgt cag atg gtttct gtt ggt acc gcg atg 2444 Tyr Met Asp Val Ser Pro Arg Gln Met Val SerVal Gly Thr Ala Met 570 575 580 att cca ttc ctg gag cac gac gat gct aaccgt gca ctg atg ggc gcg 2492 Ile Pro Phe Leu Glu His Asp Asp Ala Asn ArgAla Leu Met Gly Ala 585 590 595 aac atg cag aag cag gct gtg cca ctg attcgt gcc gag gct cct ttc 2540 Asn Met Gln Lys Gln Ala Val Pro Leu Ile ArgAla Glu Ala Pro Phe 600 605 610 gtg ggc acc ggt atg gag cag cgc gca gcatac gac gcc ggc gac ctg 2588 Val Gly Thr Gly Met Glu Gln Arg Ala Ala TyrAsp Ala Gly Asp Leu 615 620 625 gtt att acc cca gtc gca ggt gtg gtg gaaaac gtt tca gct gac ttc 2636 Val Ile Thr Pro Val Ala Gly Val Val Glu AsnVal Ser Ala Asp Phe 630 635 640 645 atc acc atc atg gct gat gac ggc aagcgc gaa acc tac ctg ctg cgt 2684 Ile Thr Ile Met Ala Asp Asp Gly Lys ArgGlu Thr Tyr Leu Leu Arg 650 655 660 aag ttc cag cgc acc aac cag ggc accagc tac aac cag aag cct ttg 2732 Lys Phe Gln Arg Thr Asn Gln Gly Thr SerTyr Asn Gln Lys Pro Leu 665 670 675 gtt aac ttg ggc gag cgc gtt gaa gctggc cag gtt att gct gat ggt 2780 Val Asn Leu Gly Glu Arg Val Glu Ala GlyGln Val Ile Ala Asp Gly 680 685 690 cca ggt acc ttc aat ggt gaa atg tccctt ggc cgt aac ctt ctg gtt 2828 Pro Gly Thr Phe Asn Gly Glu Met Ser LeuGly Arg Asn Leu Leu Val 695 700 705 gcg ttc atg cct tgg gaa ggc cac aactac gag gat gcg atc atc ctc 2876 Ala Phe Met Pro Trp Glu Gly His Asn TyrGlu Asp Ala Ile Ile Leu 710 715 720 725 aac cag aac atc gtt gag cag gacatc ttg acc tcg atc cac atc gag 2924 Asn Gln Asn Ile Val Glu Gln Asp IleLeu Thr Ser Ile His Ile Glu 730 735 740 gag cac gag atc gat gcc cgc gacact aag ctt ggc gcc gaa gaa atc 2972 Glu His Glu Ile Asp Ala Arg Asp ThrLys Leu Gly Ala Glu Glu Ile 745 750 755 acc cgc gac atc cct aat gtg tctgaa gaa gtc ctc aag gac ctc gac 3020 Thr Arg Asp Ile Pro Asn Val Ser GluGlu Val Leu Lys Asp Leu Asp 760 765 770 gac cgc ggt att gtc cgc atc ggtgct gat gtt cgt gac ggc gac atc 3068 Asp Arg Gly Ile Val Arg Ile Gly AlaAsp Val Arg Asp Gly Asp Ile 775 780 785 ctg gtc ggt aag gtc acc cct aagggc gag acc gag ctc acc ccg gaa 3116 Leu Val Gly Lys Val Thr Pro Lys GlyGlu Thr Glu Leu Thr Pro Glu 790 795 800 805 gag cgc ttg ctg cgc gca atcttc ggt gag aag gcc cgc gaa gtt cgc 3164 Glu Arg Leu Leu Arg Ala Ile PheGly Glu Lys Ala Arg Glu Val Arg 810 815 820 gat acc tcc atg aag gtg cctcac ggt gag acc ggc aag gtc atc ggc 3212 Asp Thr Ser Met Lys Val Pro HisGly Glu Thr Gly Lys Val Ile Gly 825 830 835 gtg cgt cac ttc tcc cgc gaggac gac gac gat ctg gct cct ggc gtc 3260 Val Arg His Phe Ser Arg Glu AspAsp Asp Asp Leu Ala Pro Gly Val 840 845 850 aac gag atg atc cgt atc tacgtt gct cag aag cgt aag atc cag gac 3308 Asn Glu Met Ile Arg Ile Tyr ValAla Gln Lys Arg Lys Ile Gln Asp 855 860 865 ggc gat aag ctc gct ggc cgccac ggt aac aag ggt gtt gtc ggt aaa 3356 Gly Asp Lys Leu Ala Gly Arg HisGly Asn Lys Gly Val Val Gly Lys 870 875 880 885 att ttg cct cag gaa gatatg cca ttc ctt cca gac ggc act cct gtt 3404 Ile Leu Pro Gln Glu Asp MetPro Phe Leu Pro Asp Gly Thr Pro Val 890 895 900 gac atc atc ttg aac acccac ggt gtt cca cgt cgt atg aac att ggt 3452 Asp Ile Ile Leu Asn Thr HisGly Val Pro Arg Arg Met Asn Ile Gly 905 910 915 cag gtt ctt gag acc cacctt ggc tgg ctg gca tct gct ggt tgg tcc 3500 Gln Val Leu Glu Thr His LeuGly Trp Leu Ala Ser Ala Gly Trp Ser 920 925 930 gtg gat cct gaa gat cctgag aac gct gag ctc gtc aag act ctg cct 3548 Val Asp Pro Glu Asp Pro GluAsn Ala Glu Leu Val Lys Thr Leu Pro 935 940 945 gca gac ctc ctc gag gttcct gct ggt tcc ttg act gca act cct gtg 3596 Ala Asp Leu Leu Glu Val ProAla Gly Ser Leu Thr Ala Thr Pro Val 950 955 960 965 ttc gac ggt gcg tcaaac gaa gag ctc gca ggc ctg ctc gct aat tca 3644 Phe Asp Gly Ala Ser AsnGlu Glu Leu Ala Gly Leu Leu Ala Asn Ser 970 975 980 cgt cca aac cgc gacggc gac gtc atg gtt aac gcg gat ggt aaa gca 3692 Arg Pro Asn Arg Asp GlyAsp Val Met Val Asn Ala Asp Gly Lys Ala 985 990 995 acg ctt atc gac ggtcgc tcc ggt gag cct tac ccg tac ccg gtt 3737 Thr Leu Ile Asp Gly Arg SerGly Glu Pro Tyr Pro Tyr Pro Val 1000 1005 1010 tcc atc ggc tac atg tacatg ctg aag ctg cac cac ctc gtt gac 3782 Ser Ile Gly Tyr Met Tyr Met LeuLys Leu His His Leu Val Asp 1015 1020 1025 gag aag atc cac gca cgt tccact ggt cct tac tcc atg att acc 3827 Glu Lys Ile His Ala Arg Ser Thr GlyPro Tyr Ser Met Ile Thr 1030 1035 1040 cag cag cca ctg ggt ggt aaa gcacag ttc ggt gga cag cgt ttc 3872 Gln Gln Pro Leu Gly Gly Lys Ala Gln PheGly Gly Gln Arg Phe 1045 1050 1055 ggc gaa atg gag gtg tgg gca atg caggca tac ggc gct gcc tac 3917 Gly Glu Met Glu Val Trp Ala Met Gln Ala TyrGly Ala Ala Tyr 1060 1065 1070 aca ctt cag gag ctg ctg acc atc aag tctgat gac gtg gtt ggc 3962 Thr Leu Gln Glu Leu Leu Thr Ile Lys Ser Asp AspVal Val Gly 1075 1080 1085 cgt gtc aag gtc tac gaa gca att gtg aag ggcgag aac atc ccg 4007 Arg Val Lys Val Tyr Glu Ala Ile Val Lys Gly Glu AsnIle Pro 1090 1095 1100 gat cca ggt att cct gag tcc ttc aag gtt ctc ctcaag gag ctc 4052 Asp Pro Gly Ile Pro Glu Ser Phe Lys Val Leu Leu Lys GluLeu 1105 1110 1115 cag tcc ttg tgc ctg aac gtg gag gtt ctc tcc gca gacggc act 4097 Gln Ser Leu Cys Leu Asn Val Glu Val Leu Ser Ala Asp Gly Thr1120 1125 1130 cca atg gag ctc gcg ggt gac gac gac gac ttc gat cag gcaggc 4142 Pro Met Glu Leu Ala Gly Asp Asp Asp Asp Phe Asp Gln Ala Gly1135 1140 1145 gcc tca ctt ggc atc aac ctg tcc cgt gac gag cgt tcc gacgcc 4187 Ala Ser Leu Gly Ile Asn Leu Ser Arg Asp Glu Arg Ser Asp Ala1150 1155 1160 gac acc gca tagcagatca gaaaacaacc gctagaaatc aagccataca4236 Asp Thr Ala 1165 tcccccggac attgaagaga tgttctgggg ggaaagggagttttacgtgc tcgacgtaaa 4296 cgtcttcgat gagctccgca tcggcctggc caccgccgacgacatccgcc gttggtccaa 4356 gggtgaggtc aagaagccgg agaccatcaa ctaccgaaccctcaagcctg agaaggacgg 4416 tctgttctgc gagcgtatct tcggtccaac tcgcgactgggagtgcgcct gcggtaagta 4476 caagcgtgtc cgctacaagg gcatcatctg tgaacgctgtggcgttgagg tcaccaagtc 4536 caaggtgcgc cgtgagcgca tgggacacat tgagctcgctgcaccagtaa cccacatttg 4596 gtacttcaag ggcgttccat cacgcctcgg ctaccttttggaccttgctc caaaggacct 4656 ggacctcatc atctacttcg gtgcgaacat catcaccagcgtggacgaag aggctcgcca 4716 cagcgaccag accactcttg aggcagaaat gcttctggagaagaaggacg ttgaggcaga 4776 cgcagagtct gacattgctg agcgtgctga aaagctcgaagaggatcttg ctgaacttga 4836 ggcagctggc gctaaggccg acgctcgccg caaggttcaggctgctgccg ataaggaaat 4896 gcagcacatc cgtgagcgtg cacagcgcga aatcgatcgtctcgatgagg tctggcagac 4956 cttcatcaag cttgctccaa agcagatgat ccgcgatgagaagctctacg atgaactgat 5016 cgaccgctac gaggattact tcaccggtgg tatgggtgcagagtccattg aggctttgat 5076 ccagaacttc gaccttgatg ctg 5099 2 1165 PRTCorynebacterium glutamicum 2 Val Leu Glu Gly Pro Ile Leu Ala Val Ser ArgGln Thr Lys Ser Val 1 5 10 15 Val Asp Ile Pro Gly Ala Pro Gln Arg TyrSer Phe Ala Lys Val Ser 20 25 30 Ala Pro Ile Glu Val Pro Gly Leu Leu AspLeu Gln Leu Asp Ser Tyr 35 40 45 Ser Trp Leu Ile Gly Thr Pro Glu Trp ArgAla Arg Gln Lys Glu Glu 50 55 60 Phe Gly Glu Gly Ala Arg Val Thr Ser GlyLeu Glu Asn Ile Leu Glu 65 70 75 80 Glu Leu Ser Pro Ile Gln Asp Tyr SerGly Asn Met Ser Leu Ser Leu 85 90 95 Ser Glu Pro Arg Phe Glu Asp Val LysAsn Thr Ile Asp Glu Ala Lys 100 105 110 Glu Lys Asp Ile Asn Tyr Ala AlaPro Leu Tyr Val Thr Ala Glu Phe 115 120 125 Val Asn Asn Thr Thr Gly GluIle Lys Ser Gln Thr Val Phe Ile Gly 130 135 140 Asp Phe Pro Met Met ThrAsp Lys Gly Thr Phe Ile Ile Asn Gly Thr 145 150 155 160 Glu Arg Val ValVal Ser Gln Leu Val Arg Ser Pro Gly Val Tyr Phe 165 170 175 Asp Gln ThrIle Asp Lys Ser Thr Glu Arg Pro Leu His Ala Val Lys 180 185 190 Val IlePro Ser Arg Gly Ala Trp Leu Glu Phe Asp Val Asp Lys Arg 195 200 205 AspSer Val Gly Val Arg Ile Asp Arg Lys Arg Arg Gln Pro Val Thr 210 215 220Val Leu Leu Lys Ala Leu Gly Trp Thr Thr Glu Gln Ile Thr Glu Arg 225 230235 240 Phe Gly Phe Ser Glu Ile Met Met Ser Thr Leu Glu Ser Asp Gly Val245 250 255 Ala Asn Thr Asp Glu Ala Leu Leu Glu Ile Tyr Arg Lys Gln ArgPro 260 265 270 Gly Glu Gln Pro Thr Arg Asp Leu Ala Gln Ser Leu Leu AspAsn Ser 275 280 285 Phe Phe Arg Ala Lys Arg Tyr Asp Leu Ala Arg Val GlyArg Tyr Lys 290 295 300 Ile Asn Arg Lys Leu Gly Leu Gly Gly Asp His AspGly Leu Met Thr 305 310 315 320 Leu Thr Glu Glu Asp Ile Ala Thr Thr IleGlu Tyr Leu Val Arg Leu 325 330 335 His Ala Gly Glu Arg Val Met Thr SerPro Asn Gly Glu Glu Ile Pro 340 345 350 Val Glu Thr Asp Asp Ile Asp HisPhe Gly Asn Arg Arg Leu Arg Thr 355 360 365 Val Gly Glu Leu Ile Gln AsnGln Val Arg Val Gly Leu Ser Arg Met 370 375 380 Glu Arg Val Val Arg GluArg Met Thr Thr Gln Asp Ala Glu Ser Ile 385 390 395 400 Thr Pro Thr SerLeu Ile Asn Val Arg Pro Val Ser Ala Ala Ile Arg 405 410 415 Glu Phe PheGly Thr Ser Gln Leu Ser Gln Phe Met Asp Gln Asn Asn 420 425 430 Ser LeuSer Gly Leu Thr His Lys Arg Arg Leu Ser Ala Leu Gly Pro 435 440 445 GlyGly Leu Ser Arg Glu Arg Ala Gly Ile Glu Val Arg Asp Val His 450 455 460Pro Ser His Tyr Gly Arg Met Cys Pro Ile Glu Thr Pro Glu Gly Pro 465 470475 480 Asn Ile Gly Leu Ile Gly Ser Leu Ala Ser Tyr Ala Arg Val Asn Pro485 490 495 Phe Gly Phe Ile Glu Thr Pro Tyr Arg Arg Ile Ile Asp Gly LysLeu 500 505 510 Thr Asp Gln Ile Asp Tyr Leu Thr Ala Asp Glu Glu Asp ArgPhe Val 515 520 525 Val Ala Gln Ala Asn Thr His Tyr Asp Glu Glu Gly AsnIle Thr Asp 530 535 540 Glu Thr Val Thr Val Arg Leu Lys Asp Gly Asp IleAla Met Val Gly 545 550 555 560 Arg Asn Ala Val Asp Tyr Met Asp Val SerPro Arg Gln Met Val Ser 565 570 575 Val Gly Thr Ala Met Ile Pro Phe LeuGlu His Asp Asp Ala Asn Arg 580 585 590 Ala Leu Met Gly Ala Asn Met GlnLys Gln Ala Val Pro Leu Ile Arg 595 600 605 Ala Glu Ala Pro Phe Val GlyThr Gly Met Glu Gln Arg Ala Ala Tyr 610 615 620 Asp Ala Gly Asp Leu ValIle Thr Pro Val Ala Gly Val Val Glu Asn 625 630 635 640 Val Ser Ala AspPhe Ile Thr Ile Met Ala Asp Asp Gly Lys Arg Glu 645 650 655 Thr Tyr LeuLeu Arg Lys Phe Gln Arg Thr Asn Gln Gly Thr Ser Tyr 660 665 670 Asn GlnLys Pro Leu Val Asn Leu Gly Glu Arg Val Glu Ala Gly Gln 675 680 685 ValIle Ala Asp Gly Pro Gly Thr Phe Asn Gly Glu Met Ser Leu Gly 690 695 700Arg Asn Leu Leu Val Ala Phe Met Pro Trp Glu Gly His Asn Tyr Glu 705 710715 720 Asp Ala Ile Ile Leu Asn Gln Asn Ile Val Glu Gln Asp Ile Leu Thr725 730 735 Ser Ile His Ile Glu Glu His Glu Ile Asp Ala Arg Asp Thr LysLeu 740 745 750 Gly Ala Glu Glu Ile Thr Arg Asp Ile Pro Asn Val Ser GluGlu Val 755 760 765 Leu Lys Asp Leu Asp Asp Arg Gly Ile Val Arg Ile GlyAla Asp Val 770 775 780 Arg Asp Gly Asp Ile Leu Val Gly Lys Val Thr ProLys Gly Glu Thr 785 790 795 800 Glu Leu Thr Pro Glu Glu Arg Leu Leu ArgAla Ile Phe Gly Glu Lys 805 810 815 Ala Arg Glu Val Arg Asp Thr Ser MetLys Val Pro His Gly Glu Thr 820 825 830 Gly Lys Val Ile Gly Val Arg HisPhe Ser Arg Glu Asp Asp Asp Asp 835 840 845 Leu Ala Pro Gly Val Asn GluMet Ile Arg Ile Tyr Val Ala Gln Lys 850 855 860 Arg Lys Ile Gln Asp GlyAsp Lys Leu Ala Gly Arg His Gly Asn Lys 865 870 875 880 Gly Val Val GlyLys Ile Leu Pro Gln Glu Asp Met Pro Phe Leu Pro 885 890 895 Asp Gly ThrPro Val Asp Ile Ile Leu Asn Thr His Gly Val Pro Arg 900 905 910 Arg MetAsn Ile Gly Gln Val Leu Glu Thr His Leu Gly Trp Leu Ala 915 920 925 SerAla Gly Trp Ser Val Asp Pro Glu Asp Pro Glu Asn Ala Glu Leu 930 935 940Val Lys Thr Leu Pro Ala Asp Leu Leu Glu Val Pro Ala Gly Ser Leu 945 950955 960 Thr Ala Thr Pro Val Phe Asp Gly Ala Ser Asn Glu Glu Leu Ala Gly965 970 975 Leu Leu Ala Asn Ser Arg Pro Asn Arg Asp Gly Asp Val Met ValAsn 980 985 990 Ala Asp Gly Lys Ala Thr Leu Ile Asp Gly Arg Ser Gly GluPro Tyr 995 1000 1005 Pro Tyr Pro Val Ser Ile Gly Tyr Met Tyr Met LeuLys Leu His 1010 1015 1020 His Leu Val Asp Glu Lys Ile His Ala Arg SerThr Gly Pro Tyr 1025 1030 1035 Ser Met Ile Thr Gln Gln Pro Leu Gly GlyLys Ala Gln Phe Gly 1040 1045 1050 Gly Gln Arg Phe Gly Glu Met Glu ValTrp Ala Met Gln Ala Tyr 1055 1060 1065 Gly Ala Ala Tyr Thr Leu Gln GluLeu Leu Thr Ile Lys Ser Asp 1070 1075 1080 Asp Val Val Gly Arg Val LysVal Tyr Glu Ala Ile Val Lys Gly 1085 1090 1095 Glu Asn Ile Pro Asp ProGly Ile Pro Glu Ser Phe Lys Val Leu 1100 1105 1110 Leu Lys Glu Leu GlnSer Leu Cys Leu Asn Val Glu Val Leu Ser 1115 1120 1125 Ala Asp Gly ThrPro Met Glu Leu Ala Gly Asp Asp Asp Asp Phe 1130 1135 1140 Asp Gln AlaGly Ala Ser Leu Gly Ile Asn Leu Ser Arg Asp Glu 1145 1150 1155 Arg SerAsp Ala Asp Thr Ala 1160 1165 3 5099 DNA Corynebacterium glutamicum CDS(702)..(4196) 3 acaatgtgac tcgtgatttt tgggtggatc agcgtaccgg tttggttgtcgatctagctg 60 aaaatattga tgatttttac ggcgaccgca gcggccagaa gtacgaacagaaattgcttt 120 tcgacgcctc cctcgacgat gcagctgtct ctaagctggt tgcacaggccgaaagcatcc 180 ctgatggaga tgtgagcaaa atcgcaaata ccgtaggtat tgtgatcggtgcggtattgg 240 ctctcgtggg cctggccggg tgttttgggg cgtttgggaa gaaacgtcgagaagcttaac 300 ctgctgttca aatagatttt ccctgtttcg aattgcggaa accccgggtttgtttgctag 360 ggtgcctcgt agaaggggtc aagaagattt ctgggaaacg cgcccgtgcggttggttgct 420 aatagcacgc ggagcaccag atgaaaaatc tcccctttac tttcgcgcgcgattggtata 480 ctctgagtcg ttgcgttgga attcgtgact ctttttcgtt cctgtagcgccaagaccttg 540 atcaaggtgg tttaaaaaaa ccgatttgac aaggtcattc agtgctatctggagtcgttc 600 agggggatcg ggttcctcag cagaccaatt gctcaaaaat accagcggtgttgatctgca 660 cttaatggcc ttgaccagcc aggtgcaatt acccgcgtga g gtg ctg gaagga ctc 716 Val Leu Glu Gly Leu 1 5 atc ttg gca gtc tcc cgc cag acc aagtca gtc gtc gat att ccc ggt 764 Ile Leu Ala Val Ser Arg Gln Thr Lys SerVal Val Asp Ile Pro Gly 10 15 20 gca ccg cag cgt tat tct ttc gcg aag gtgtcc gca ccc att gag gtg 812 Ala Pro Gln Arg Tyr Ser Phe Ala Lys Val SerAla Pro Ile Glu Val 25 30 35 ccc ggg cta cta gat ctt caa ctg gat tct tactcc tgg ctg att ggt 860 Pro Gly Leu Leu Asp Leu Gln Leu Asp Ser Tyr SerTrp Leu Ile Gly 40 45 50 acg cct gag tgg cgt gct cgt cag aag gaa gaa ttcggc gag gga gcc 908 Thr Pro Glu Trp Arg Ala Arg Gln Lys Glu Glu Phe GlyGlu Gly Ala 55 60 65 cgc gta acc agc ggc ctt gag aac att ctc gag gag ctctcc cca atc 956 Arg Val Thr Ser Gly Leu Glu Asn Ile Leu Glu Glu Leu SerPro Ile 70 75 80 85 cag gat tac tct gga aac atg tcc ctg agc ctt tcg gagcca cgc ttc 1004 Gln Asp Tyr Ser Gly Asn Met Ser Leu Ser Leu Ser Glu ProArg Phe 90 95 100 gaa gac gtc aag aac acc att gac gag gcg aaa gaa aaggac atc aac 1052 Glu Asp Val Lys Asn Thr Ile Asp Glu Ala Lys Glu Lys AspIle Asn 105 110 115 tac gcg gcg cca ctg tat gtg acc gcg gag ttc gtc aacaac acc acc 1100 Tyr Ala Ala Pro Leu Tyr Val Thr Ala Glu Phe Val Asn AsnThr Thr 120 125 130 ggt gaa atc aag tct cag act gtc ttc atc ggc gat ttccca atg atg 1148 Gly Glu Ile Lys Ser Gln Thr Val Phe Ile Gly Asp Phe ProMet Met 135 140 145 acg gac aag gga acg ttc atc atc aac gga acc gaa cgcgtt gtg gtc 1196 Thr Asp Lys Gly Thr Phe Ile Ile Asn Gly Thr Glu Arg ValVal Val 150 155 160 165 agc cag ctc gtc cgc tcc ccg ggc gtg tac ttt gaccag acc atc gat 1244 Ser Gln Leu Val Arg Ser Pro Gly Val Tyr Phe Asp GlnThr Ile Asp 170 175 180 aag tca act gag cgt cca ctg cac gcc gtg aag gttatt cct ttc cgt 1292 Lys Ser Thr Glu Arg Pro Leu His Ala Val Lys Val IlePro Phe Arg 185 190 195 ggt gct tgg ctt gag ttt gac gtc gat aag cgc gattcg gtt ggt gtt 1340 Gly Ala Trp Leu Glu Phe Asp Val Asp Lys Arg Asp SerVal Gly Val 200 205 210 cgt att gac cgc aag cgt cgc cag cca gtc acc gtactg ctg aag gct 1388 Arg Ile Asp Arg Lys Arg Arg Gln Pro Val Thr Val LeuLeu Lys Ala 215 220 225 ctt ggc tgg acc act gag cag atc acc gag cgt ttcggt ttc tct gaa 1436 Leu Gly Trp Thr Thr Glu Gln Ile Thr Glu Arg Phe GlyPhe Ser Glu 230 235 240 245 atc atg atg tcc acc ctc gag tcc gat ggt gtagca aac acc gat gag 1484 Ile Met Met Ser Thr Leu Glu Ser Asp Gly Val AlaAsn Thr Asp Glu 250 255 260 gca ttg ctg gag atc tac cgc aag cag cgt ccaggc gag cag cct acc 1532 Ala Leu Leu Glu Ile Tyr Arg Lys Gln Arg Pro GlyGlu Gln Pro Thr 265 270 275 cgc gac ctt gcg cag tcc ctc ctg gac aac agcttc ttc cgt gca aag 1580 Arg Asp Leu Ala Gln Ser Leu Leu Asp Asn Ser PhePhe Arg Ala Lys 280 285 290 cgc tac gac ctg gct cgc gtt ggt cgt tac aagatc aac cgc aag ctc 1628 Arg Tyr Asp Leu Ala Arg Val Gly Arg Tyr Lys IleAsn Arg Lys Leu 295 300 305 ggc ctt ggt ggc gac cac gat ggt ttg atg actctt act gaa gag gac 1676 Gly Leu Gly Gly Asp His Asp Gly Leu Met Thr LeuThr Glu Glu Asp 310 315 320 325 atc gca acc acc atc gag tac ctg gtg cgtctg cac gca ggt gag cgc 1724 Ile Ala Thr Thr Ile Glu Tyr Leu Val Arg LeuHis Ala Gly Glu Arg 330 335 340 gtc atg act tct cca aat ggt gaa gag atccca gtc gag acc gat gac 1772 Val Met Thr Ser Pro Asn Gly Glu Glu Ile ProVal Glu Thr Asp Asp 345 350 355 atc gac cac ttt ggt aac cgt cgt ctg cgtacc gtt ggc gaa ctg atc 1820 Ile Asp His Phe Gly Asn Arg Arg Leu Arg ThrVal Gly Glu Leu Ile 360 365 370 cag aac cag gtc cgt gtc ggc ctg tcc cgcatg gag cgc gtt gtt cgt 1868 Gln Asn Gln Val Arg Val Gly Leu Ser Arg MetGlu Arg Val Val Arg 375 380 385 gag cgt atg acc acc cag gat gcg gag tccatt act cct act tcc ttg 1916 Glu Arg Met Thr Thr Gln Asp Ala Glu Ser IleThr Pro Thr Ser Leu 390 395 400 405 atc aac gtt cgt cct gtc tct gca gctatc cgt gag ttc ttc gga act 1964 Ile Asn Val Arg Pro Val Ser Ala Ala IleArg Glu Phe Phe Gly Thr 410 415 420 tcc cag ctg tct cag ttc atg gtc cagaac aac tcc ctg tct ggt ttg 2012 Ser Gln Leu Ser Gln Phe Met Val Gln AsnAsn Ser Leu Ser Gly Leu 425 430 435 act cac aag cgt cgt ctg tcg gct ctgggc ccg ggt ggt ctg tcc cgt 2060 Thr His Lys Arg Arg Leu Ser Ala Leu GlyPro Gly Gly Leu Ser Arg 440 445 450 gag cgc gcc ggc atc gag gtt cga gacgtt cac cca tct cac tac ggc 2108 Glu Arg Ala Gly Ile Glu Val Arg Asp ValHis Pro Ser His Tyr Gly 455 460 465 cgt atg tgc cca att gag act ccg gaaggt cca aac att ggc ctg atc 2156 Arg Met Cys Pro Ile Glu Thr Pro Glu GlyPro Asn Ile Gly Leu Ile 470 475 480 485 ggt tcc ttg gct tcc tat gct cgagtg aac cca ttc ggt ttc att gag 2204 Gly Ser Leu Ala Ser Tyr Ala Arg ValAsn Pro Phe Gly Phe Ile Glu 490 495 500 acc cca tac cgt cgc atc atc gacggc aag ctg acc gac cag att gac 2252 Thr Pro Tyr Arg Arg Ile Ile Asp GlyLys Leu Thr Asp Gln Ile Asp 505 510 515 tac ctt acc gct gat gag gaa gaccgc ttc gtt gtt gcg cag gca aac 2300 Tyr Leu Thr Ala Asp Glu Glu Asp ArgPhe Val Val Ala Gln Ala Asn 520 525 530 acg cac tac gac gaa gag ggc aacatc acc gat gag acc gtc act gtt 2348 Thr His Tyr Asp Glu Glu Gly Asn IleThr Asp Glu Thr Val Thr Val 535 540 545 cgt ctg aag gac ggc gac atc gccatg gtt ggc cgc aac gcg gtt gat 2396 Arg Leu Lys Asp Gly Asp Ile Ala MetVal Gly Arg Asn Ala Val Asp 550 555 560 565 tac atg gac gtt tcc cct cgtcag atg gtt tct gtt ggt acc gcg atg 2444 Tyr Met Asp Val Ser Pro Arg GlnMet Val Ser Val Gly Thr Ala Met 570 575 580 att cca ttc ctg gag cac gacgat gct aac cgt gca ctg atg ggc gcg 2492 Ile Pro Phe Leu Glu His Asp AspAla Asn Arg Ala Leu Met Gly Ala 585 590 595 aac atg cag aag cag gct gtgcca ctg att cgt gcc gag gct cct ttc 2540 Asn Met Gln Lys Gln Ala Val ProLeu Ile Arg Ala Glu Ala Pro Phe 600 605 610 gtg ggc acc ggt atg gag cagcgc gca gca tac gac gcc ggc gac ctg 2588 Val Gly Thr Gly Met Glu Gln ArgAla Ala Tyr Asp Ala Gly Asp Leu 615 620 625 gtt att acc cca gtc gca ggtgtg gtg gaa aac gtt tca gct gac ttc 2636 Val Ile Thr Pro Val Ala Gly ValVal Glu Asn Val Ser Ala Asp Phe 630 635 640 645 atc acc atc atg gct gatgac ggc aag cgc gaa acc tac ctg ctg cgt 2684 Ile Thr Ile Met Ala Asp AspGly Lys Arg Glu Thr Tyr Leu Leu Arg 650 655 660 aag ttc cag cgc acc aaccag ggc acc agc tac aac cag aag cct ttg 2732 Lys Phe Gln Arg Thr Asn GlnGly Thr Ser Tyr Asn Gln Lys Pro Leu 665 670 675 gtt aac ttg ggc gag cgcgtt gaa gct ggc cag gtt att gct gat ggt 2780 Val Asn Leu Gly Glu Arg ValGlu Ala Gly Gln Val Ile Ala Asp Gly 680 685 690 cca ggt acc ttc aat ggtgaa atg tcc ctt ggc cgt aac ctt ctg gtt 2828 Pro Gly Thr Phe Asn Gly GluMet Ser Leu Gly Arg Asn Leu Leu Val 695 700 705 gcg ttc atg cct tgg gaaggc cac aac tac gag gat gcg atc atc ctc 2876 Ala Phe Met Pro Trp Glu GlyHis Asn Tyr Glu Asp Ala Ile Ile Leu 710 715 720 725 aac cag aac atc gttgag cag gac atc ttg acc tcg atc cac atc gag 2924 Asn Gln Asn Ile Val GluGln Asp Ile Leu Thr Ser Ile His Ile Glu 730 735 740 gag cac gag atc gatgcc cgc gac act aag ctt ggc gcc gaa gaa atc 2972 Glu His Glu Ile Asp AlaArg Asp Thr Lys Leu Gly Ala Glu Glu Ile 745 750 755 acc cgc gac atc cctaat gtg tct gaa gaa gtc ctc aag gac ctc gac 3020 Thr Arg Asp Ile Pro AsnVal Ser Glu Glu Val Leu Lys Asp Leu Asp 760 765 770 gac cgc ggt att gtccgc atc ggt gct gat gtt cgt gac ggc gac atc 3068 Asp Arg Gly Ile Val ArgIle Gly Ala Asp Val Arg Asp Gly Asp Ile 775 780 785 ctg gtc ggt aag gtcacc cct aag ggc gag acc gag ctc acc ccg gaa 3116 Leu Val Gly Lys Val ThrPro Lys Gly Glu Thr Glu Leu Thr Pro Glu 790 795 800 805 gag cgc ttg ctgcgc gca atc ttc ggt gag aag gcc cgc gaa gtt cgc 3164 Glu Arg Leu Leu ArgAla Ile Phe Gly Glu Lys Ala Arg Glu Val Arg 810 815 820 gat acc tcc atgaag gtg cct cac ggt gag acc ggc aag gtc atc ggc 3212 Asp Thr Ser Met LysVal Pro His Gly Glu Thr Gly Lys Val Ile Gly 825 830 835 gtg cgt cac ttctcc cgc gag gac gac gac gat ctg gct cct ggc gtc 3260 Val Arg His Phe SerArg Glu Asp Asp Asp Asp Leu Ala Pro Gly Val 840 845 850 aac gag atg atccgt atc tac gtt gct cag aag cgt aag atc cag gac 3308 Asn Glu Met Ile ArgIle Tyr Val Ala Gln Lys Arg Lys Ile Gln Asp 855 860 865 ggc gat aag ctcgct ggc cgc cac ggt aac aag ggt gtt gtc ggt aaa 3356 Gly Asp Lys Leu AlaGly Arg His Gly Asn Lys Gly Val Val Gly Lys 870 875 880 885 att ttg cctcag gaa gat atg cca ttc ctt cca gac ggc act cct gtt 3404 Ile Leu Pro GlnGlu Asp Met Pro Phe Leu Pro Asp Gly Thr Pro Val 890 895 900 gac atc atcttg aac acc cac ggt gtt cca cgt cgt atg aac att ggt 3452 Asp Ile Ile LeuAsn Thr His Gly Val Pro Arg Arg Met Asn Ile Gly 905 910 915 cag gtt cttgag acc cac ctt ggc tgg ctg gca tct gct ggt tgg tcc 3500 Gln Val Leu GluThr His Leu Gly Trp Leu Ala Ser Ala Gly Trp Ser 920 925 930 gtg gat cctgaa gat cct gag aac gct gag ctc gtc aag act ctg cct 3548 Val Asp Pro GluAsp Pro Glu Asn Ala Glu Leu Val Lys Thr Leu Pro 935 940 945 gca gac ctcctc gag gtt cct gct ggt tcc ttg act gca act cct gtg 3596 Ala Asp Leu LeuGlu Val Pro Ala Gly Ser Leu Thr Ala Thr Pro Val 950 955 960 965 ttc gacggt gcg tca aac gaa gag ctc gca ggc ctg ctc gct aat tca 3644 Phe Asp GlyAla Ser Asn Glu Glu Leu Ala Gly Leu Leu Ala Asn Ser 970 975 980 cgt ccaaac cgc gac ggc gac gtc atg gtt aac gcg gat ggt aaa gca 3692 Arg Pro AsnArg Asp Gly Asp Val Met Val Asn Ala Asp Gly Lys Ala 985 990 995 acg cttatc gac ggt cgc tcc ggt gag cct tac ccg tac ccg gtt 3737 Thr Leu Ile AspGly Arg Ser Gly Glu Pro Tyr Pro Tyr Pro Val 1000 1005 1010 tcc atc ggctac atg tac atg ctg aag ctg cac cac ctc gtt gac 3782 Ser Ile Gly Tyr MetTyr Met Leu Lys Leu His His Leu Val Asp 1015 1020 1025 gag aag atc cacgca cgt tcc act ggt cct tac tcc atg att acc 3827 Glu Lys Ile His Ala ArgSer Thr Gly Pro Tyr Ser Met Ile Thr 1030 1035 1040 cag cag cca ctg ggtggt aaa gca cag ttc ggt gga cag cgt ttc 3872 Gln Gln Pro Leu Gly Gly LysAla Gln Phe Gly Gly Gln Arg Phe 1045 1050 1055 ggc gaa atg gag gtg tgggca atg cag gca tac ggc gct gcc tac 3917 Gly Glu Met Glu Val Trp Ala MetGln Ala Tyr Gly Ala Ala Tyr 1060 1065 1070 aca ctt cag gag ctg ctg accatc aag tct gat gac gtg gtt ggc 3962 Thr Leu Gln Glu Leu Leu Thr Ile LysSer Asp Asp Val Val Gly 1075 1080 1085 cgt gtc aag gtc tac gaa gca attgtg aag ggc gag aac atc ccg 4007 Arg Val Lys Val Tyr Glu Ala Ile Val LysGly Glu Asn Ile Pro 1090 1095 1100 gat cca ggt att cct gag tcc ttc aaggtt ctc ctc aag gag ctc 4052 Asp Pro Gly Ile Pro Glu Ser Phe Lys Val LeuLeu Lys Glu Leu 1105 1110 1115 cag tcc ttg tgc ctg aac gtg gag gtt ctctcc gca gac ggc act 4097 Gln Ser Leu Cys Leu Asn Val Glu Val Leu Ser AlaAsp Gly Thr 1120 1125 1130 cca atg gag ctc gcg ggt gac gac gac gac ttcgat cag gca ggc 4142 Pro Met Glu Leu Ala Gly Asp Asp Asp Asp Phe Asp GlnAla Gly 1135 1140 1145 gcc tca ctt ggc atc aac ctg tcc cgt gac gag cgttcc gac gcc 4187 Ala Ser Leu Gly Ile Asn Leu Ser Arg Asp Glu Arg Ser AspAla 1150 1155 1160 gac acc gca tagcagatca gaaaacaacc gctagaaatcaagccataca 4236 Asp Thr Ala 1165 tcccccggac attgaagaga tgttctggggggaaagggag ttttacgtgc tcgacgtaaa 4296 cgtcttcgat gagctccgca tcggcctggccaccgccgac gacatccgcc gttggtccaa 4356 gggtgaggtc aagaagccgg agaccatcaactaccgaacc ctcaagcctg agaaggacgg 4416 tctgttctgc gagcgtatct tcggtccaactcgcgactgg gagtgcgcct gcggtaagta 4476 caagcgtgtc cgctacaagg gcatcatctgtgaacgctgt ggcgttgagg tcaccaagtc 4536 caaggtgcgc cgtgagcgca tgggacacattgagctcgct gcaccagtaa cccacatttg 4596 gtacttcaag ggcgttccat cacgcctcggctaccttttg gaccttgctc caaaggacct 4656 ggacctcatc atctacttcg gtgcgaacatcatcaccagc gtggacgaag aggctcgcca 4716 cagcgaccag accactcttg aggcagaaatgcttctggag aagaaggacg ttgaggcaga 4776 cgcagagtct gacattgctg agcgtgctgaaaagctcgaa gaggatcttg ctgaacttga 4836 ggcagctggc gctaaggccg acgctcgccgcaaggttcag gctgctgccg ataaggaaat 4896 gcagcacatc cgtgagcgtg cacagcgcgaaatcgatcgt ctcgatgagg tctggcagac 4956 cttcatcaag cttgctccaa agcagatgatccgcgatgag aagctctacg atgaactgat 5016 cgaccgctac gaggattact tcaccggtggtatgggtgca gagtccattg aggctttgat 5076 ccagaacttc gaccttgatg ctg 5099 41165 PRT Corynebacterium glutamicum 4 Val Leu Glu Gly Leu Ile Leu AlaVal Ser Arg Gln Thr Lys Ser Val 1 5 10 15 Val Asp Ile Pro Gly Ala ProGln Arg Tyr Ser Phe Ala Lys Val Ser 20 25 30 Ala Pro Ile Glu Val Pro GlyLeu Leu Asp Leu Gln Leu Asp Ser Tyr 35 40 45 Ser Trp Leu Ile Gly Thr ProGlu Trp Arg Ala Arg Gln Lys Glu Glu 50 55 60 Phe Gly Glu Gly Ala Arg ValThr Ser Gly Leu Glu Asn Ile Leu Glu 65 70 75 80 Glu Leu Ser Pro Ile GlnAsp Tyr Ser Gly Asn Met Ser Leu Ser Leu 85 90 95 Ser Glu Pro Arg Phe GluAsp Val Lys Asn Thr Ile Asp Glu Ala Lys 100 105 110 Glu Lys Asp Ile AsnTyr Ala Ala Pro Leu Tyr Val Thr Ala Glu Phe 115 120 125 Val Asn Asn ThrThr Gly Glu Ile Lys Ser Gln Thr Val Phe Ile Gly 130 135 140 Asp Phe ProMet Met Thr Asp Lys Gly Thr Phe Ile Ile Asn Gly Thr 145 150 155 160 GluArg Val Val Val Ser Gln Leu Val Arg Ser Pro Gly Val Tyr Phe 165 170 175Asp Gln Thr Ile Asp Lys Ser Thr Glu Arg Pro Leu His Ala Val Lys 180 185190 Val Ile Pro Phe Arg Gly Ala Trp Leu Glu Phe Asp Val Asp Lys Arg 195200 205 Asp Ser Val Gly Val Arg Ile Asp Arg Lys Arg Arg Gln Pro Val Thr210 215 220 Val Leu Leu Lys Ala Leu Gly Trp Thr Thr Glu Gln Ile Thr GluArg 225 230 235 240 Phe Gly Phe Ser Glu Ile Met Met Ser Thr Leu Glu SerAsp Gly Val 245 250 255 Ala Asn Thr Asp Glu Ala Leu Leu Glu Ile Tyr ArgLys Gln Arg Pro 260 265 270 Gly Glu Gln Pro Thr Arg Asp Leu Ala Gln SerLeu Leu Asp Asn Ser 275 280 285 Phe Phe Arg Ala Lys Arg Tyr Asp Leu AlaArg Val Gly Arg Tyr Lys 290 295 300 Ile Asn Arg Lys Leu Gly Leu Gly GlyAsp His Asp Gly Leu Met Thr 305 310 315 320 Leu Thr Glu Glu Asp Ile AlaThr Thr Ile Glu Tyr Leu Val Arg Leu 325 330 335 His Ala Gly Glu Arg ValMet Thr Ser Pro Asn Gly Glu Glu Ile Pro 340 345 350 Val Glu Thr Asp AspIle Asp His Phe Gly Asn Arg Arg Leu Arg Thr 355 360 365 Val Gly Glu LeuIle Gln Asn Gln Val Arg Val Gly Leu Ser Arg Met 370 375 380 Glu Arg ValVal Arg Glu Arg Met Thr Thr Gln Asp Ala Glu Ser Ile 385 390 395 400 ThrPro Thr Ser Leu Ile Asn Val Arg Pro Val Ser Ala Ala Ile Arg 405 410 415Glu Phe Phe Gly Thr Ser Gln Leu Ser Gln Phe Met Val Gln Asn Asn 420 425430 Ser Leu Ser Gly Leu Thr His Lys Arg Arg Leu Ser Ala Leu Gly Pro 435440 445 Gly Gly Leu Ser Arg Glu Arg Ala Gly Ile Glu Val Arg Asp Val His450 455 460 Pro Ser His Tyr Gly Arg Met Cys Pro Ile Glu Thr Pro Glu GlyPro 465 470 475 480 Asn Ile Gly Leu Ile Gly Ser Leu Ala Ser Tyr Ala ArgVal Asn Pro 485 490 495 Phe Gly Phe Ile Glu Thr Pro Tyr Arg Arg Ile IleAsp Gly Lys Leu 500 505 510 Thr Asp Gln Ile Asp Tyr Leu Thr Ala Asp GluGlu Asp Arg Phe Val 515 520 525 Val Ala Gln Ala Asn Thr His Tyr Asp GluGlu Gly Asn Ile Thr Asp 530 535 540 Glu Thr Val Thr Val Arg Leu Lys AspGly Asp Ile Ala Met Val Gly 545 550 555 560 Arg Asn Ala Val Asp Tyr MetAsp Val Ser Pro Arg Gln Met Val Ser 565 570 575 Val Gly Thr Ala Met IlePro Phe Leu Glu His Asp Asp Ala Asn Arg 580 585 590 Ala Leu Met Gly AlaAsn Met Gln Lys Gln Ala Val Pro Leu Ile Arg 595 600 605 Ala Glu Ala ProPhe Val Gly Thr Gly Met Glu Gln Arg Ala Ala Tyr 610 615 620 Asp Ala GlyAsp Leu Val Ile Thr Pro Val Ala Gly Val Val Glu Asn 625 630 635 640 ValSer Ala Asp Phe Ile Thr Ile Met Ala Asp Asp Gly Lys Arg Glu 645 650 655Thr Tyr Leu Leu Arg Lys Phe Gln Arg Thr Asn Gln Gly Thr Ser Tyr 660 665670 Asn Gln Lys Pro Leu Val Asn Leu Gly Glu Arg Val Glu Ala Gly Gln 675680 685 Val Ile Ala Asp Gly Pro Gly Thr Phe Asn Gly Glu Met Ser Leu Gly690 695 700 Arg Asn Leu Leu Val Ala Phe Met Pro Trp Glu Gly His Asn TyrGlu 705 710 715 720 Asp Ala Ile Ile Leu Asn Gln Asn Ile Val Glu Gln AspIle Leu Thr 725 730 735 Ser Ile His Ile Glu Glu His Glu Ile Asp Ala ArgAsp Thr Lys Leu 740 745 750 Gly Ala Glu Glu Ile Thr Arg Asp Ile Pro AsnVal Ser Glu Glu Val 755 760 765 Leu Lys Asp Leu Asp Asp Arg Gly Ile ValArg Ile Gly Ala Asp Val 770 775 780 Arg Asp Gly Asp Ile Leu Val Gly LysVal Thr Pro Lys Gly Glu Thr 785 790 795 800 Glu Leu Thr Pro Glu Glu ArgLeu Leu Arg Ala Ile Phe Gly Glu Lys 805 810 815 Ala Arg Glu Val Arg AspThr Ser Met Lys Val Pro His Gly Glu Thr 820 825 830 Gly Lys Val Ile GlyVal Arg His Phe Ser Arg Glu Asp Asp Asp Asp 835 840 845 Leu Ala Pro GlyVal Asn Glu Met Ile Arg Ile Tyr Val Ala Gln Lys 850 855 860 Arg Lys IleGln Asp Gly Asp Lys Leu Ala Gly Arg His Gly Asn Lys 865 870 875 880 GlyVal Val Gly Lys Ile Leu Pro Gln Glu Asp Met Pro Phe Leu Pro 885 890 895Asp Gly Thr Pro Val Asp Ile Ile Leu Asn Thr His Gly Val Pro Arg 900 905910 Arg Met Asn Ile Gly Gln Val Leu Glu Thr His Leu Gly Trp Leu Ala 915920 925 Ser Ala Gly Trp Ser Val Asp Pro Glu Asp Pro Glu Asn Ala Glu Leu930 935 940 Val Lys Thr Leu Pro Ala Asp Leu Leu Glu Val Pro Ala Gly SerLeu 945 950 955 960 Thr Ala Thr Pro Val Phe Asp Gly Ala Ser Asn Glu GluLeu Ala Gly 965 970 975 Leu Leu Ala Asn Ser Arg Pro Asn Arg Asp Gly AspVal Met Val Asn 980 985 990 Ala Asp Gly Lys Ala Thr Leu Ile Asp Gly ArgSer Gly Glu Pro Tyr 995 1000 1005 Pro Tyr Pro Val Ser Ile Gly Tyr MetTyr Met Leu Lys Leu His 1010 1015 1020 His Leu Val Asp Glu Lys Ile HisAla Arg Ser Thr Gly Pro Tyr 1025 1030 1035 Ser Met Ile Thr Gln Gln ProLeu Gly Gly Lys Ala Gln Phe Gly 1040 1045 1050 Gly Gln Arg Phe Gly GluMet Glu Val Trp Ala Met Gln Ala Tyr 1055 1060 1065 Gly Ala Ala Tyr ThrLeu Gln Glu Leu Leu Thr Ile Lys Ser Asp 1070 1075 1080 Asp Val Val GlyArg Val Lys Val Tyr Glu Ala Ile Val Lys Gly 1085 1090 1095 Glu Asn IlePro Asp Pro Gly Ile Pro Glu Ser Phe Lys Val Leu 1100 1105 1110 Leu LysGlu Leu Gln Ser Leu Cys Leu Asn Val Glu Val Leu Ser 1115 1120 1125 AlaAsp Gly Thr Pro Met Glu Leu Ala Gly Asp Asp Asp Asp Phe 1130 1135 1140Asp Gln Ala Gly Ala Ser Leu Gly Ile Asn Leu Ser Arg Asp Glu 1145 11501155 Arg Ser Asp Ala Asp Thr Ala 1160 1165 5 5099 DNA Corynebacteriumglutamicum CDS (702)..(4196) 5 acaatgtgac tcgtgatttt tgggtggatcagcgtaccgg tttggttgtc gatctagctg 60 aaaatattga tgatttttac ggcgaccgcagcggccagaa gtacgaacag aaattgcttt 120 tcgacgcctc cctcgacgat gcagctgtctctaagctggt tgcacaggcc gaaagcatcc 180 ctgatggaga tgtgagcaaa atcgcaaataccgtaggtat tgtgatcggt gcggtattgg 240 ctctcgtggg cctggccggg tgttttggggcgtttgggaa gaaacgtcga gaagcttaac 300 ctgctgttca aatagatttt ccctgtttcgaattgcggaa accccgggtt tgtttgctag 360 ggtgcctcgt agaaggggtc aagaagatttctgggaaacg cgcccgtgcg gttggttgct 420 aatagcacgc ggagcaccag atgaaaaatctcccctttac tttcgcgcgc gattggtata 480 ctctgagtcg ttgcgttgga attcgtgactctttttcgtt cctgtagcgc caagaccttg 540 atcaaggtgg tttaaaaaaa ccgatttgacaaggtcattc agtgctatct ggagtcgttc 600 agggggatcg ggttcctcag cagaccaattgctcaaaaat accagcggtg ttgatctgca 660 cttaatggcc ttgaccagcc aggtgcaattacccgcgtga g gtg ctg gaa gga ccc 716 Val Leu Glu Gly Pro 1 5 atc ttg gcagtc tcc cgc cag acc aag tca gtc gtc gat att ccc ggt 764 Ile Leu Ala ValSer Arg Gln Thr Lys Ser Val Val Asp Ile Pro Gly 10 15 20 gca ccg cag cgttat tct ttc gcg aag gtg tcc gca ccc att gag gtg 812 Ala Pro Gln Arg TyrSer Phe Ala Lys Val Ser Ala Pro Ile Glu Val 25 30 35 ccc ggg cta cta gatctt caa ctg gat tct tac tcc tgg ctg att ggt 860 Pro Gly Leu Leu Asp LeuGln Leu Asp Ser Tyr Ser Trp Leu Ile Gly 40 45 50 acg cct gag tgg cgt gctcgt cag aag gaa gaa ttc ggc gag gga gcc 908 Thr Pro Glu Trp Arg Ala ArgGln Lys Glu Glu Phe Gly Glu Gly Ala 55 60 65 cgc gta acc agc ggc ctt gagaac att ctc gag gag ctc tcc cca atc 956 Arg Val Thr Ser Gly Leu Glu AsnIle Leu Glu Glu Leu Ser Pro Ile 70 75 80 85 cag gat tac tct gga aac atgtcc ctg agc ctt tcg gag cca cgc ttc 1004 Gln Asp Tyr Ser Gly Asn Met SerLeu Ser Leu Ser Glu Pro Arg Phe 90 95 100 gaa gac gtc aag aac acc attgac gag gcg aaa gaa aag gac atc aac 1052 Glu Asp Val Lys Asn Thr Ile AspGlu Ala Lys Glu Lys Asp Ile Asn 105 110 115 tac gcg gcg cca ctg tat gtgacc gcg gag ttc gtc aac aac acc acc 1100 Tyr Ala Ala Pro Leu Tyr Val ThrAla Glu Phe Val Asn Asn Thr Thr 120 125 130 ggt gaa atc aag tct cag actgtc ttc atc ggc gat ttc cca atg atg 1148 Gly Glu Ile Lys Ser Gln Thr ValPhe Ile Gly Asp Phe Pro Met Met 135 140 145 acg gac aag gga acg ttc atcatc aac gga acc gaa cgc gtt gtg gtc 1196 Thr Asp Lys Gly Thr Phe Ile IleAsn Gly Thr Glu Arg Val Val Val 150 155 160 165 agc cag ctc gtc cgc tccccg ggc gtg tac ttt gac cag acc atc gat 1244 Ser Gln Leu Val Arg Ser ProGly Val Tyr Phe Asp Gln Thr Ile Asp 170 175 180 aag tca act gag cgt ccactg cac gcc gtg aag gtt att cct tcc cgt 1292 Lys Ser Thr Glu Arg Pro LeuHis Ala Val Lys Val Ile Pro Ser Arg 185 190 195 ggt gct tgg ctt gag tttgac gtc gat aag cgc gat tcg gtt ggt gtt 1340 Gly Ala Trp Leu Glu Phe AspVal Asp Lys Arg Asp Ser Val Gly Val 200 205 210 cgt att gac cgc aag cgtcgc cag cca gtc acc gta ctg ctg aag gct 1388 Arg Ile Asp Arg Lys Arg ArgGln Pro Val Thr Val Leu Leu Lys Ala 215 220 225 ctt ggc tgg acc act gagcag atc acc gag cgt ttc ggt ttc tct gaa 1436 Leu Gly Trp Thr Thr Glu GlnIle Thr Glu Arg Phe Gly Phe Ser Glu 230 235 240 245 atc atg atg tcc accctc gag tcc gat ggt gta gca aac acc gat gag 1484 Ile Met Met Ser Thr LeuGlu Ser Asp Gly Val Ala Asn Thr Asp Glu 250 255 260 gca ttg ctg gag atctac cgc aag cag cgt cca ggc gag cag cct acc 1532 Ala Leu Leu Glu Ile TyrArg Lys Gln Arg Pro Gly Glu Gln Pro Thr 265 270 275 cgc gac ctt gcg cagtcc ctc ctg gac aac agc ttc ttc cgt gca aag 1580 Arg Asp Leu Ala Gln SerLeu Leu Asp Asn Ser Phe Phe Arg Ala Lys 280 285 290 cgc tac gac ctg gctcgc gtt ggt cgt tac aag atc aac cgc aag ctc 1628 Arg Tyr Asp Leu Ala ArgVal Gly Arg Tyr Lys Ile Asn Arg Lys Leu 295 300 305 ggc ctt ggt ggc gaccac gat ggt ttg atg act ctt act gaa gag gac 1676 Gly Leu Gly Gly Asp HisAsp Gly Leu Met Thr Leu Thr Glu Glu Asp 310 315 320 325 atc gca acc accatc gag tac ctg gtg cgt ctg cac gca ggt gag cgc 1724 Ile Ala Thr Thr IleGlu Tyr Leu Val Arg Leu His Ala Gly Glu Arg 330 335 340 gtc atg act tctcca aat ggt gaa gag atc cca gtc gag acc gat gac 1772 Val Met Thr Ser ProAsn Gly Glu Glu Ile Pro Val Glu Thr Asp Asp 345 350 355 atc gac cac tttggt aac cgt cgt ctg cgt acc gtt ggc gaa ctg atc 1820 Ile Asp His Phe GlyAsn Arg Arg Leu Arg Thr Val Gly Glu Leu Ile 360 365 370 cag aac cag gtccgt gtc ggc ctg tcc cgc atg gag cgc gtt gtt cgt 1868 Gln Asn Gln Val ArgVal Gly Leu Ser Arg Met Glu Arg Val Val Arg 375 380 385 gag cgt atg accacc cag gat gcg gag tcc att act cct act tcc ttg 1916 Glu Arg Met Thr ThrGln Asp Ala Glu Ser Ile Thr Pro Thr Ser Leu 390 395 400 405 atc aac gttcgt cct gtc tct gca gct atc cgt gag ttc ttc gga act 1964 Ile Asn Val ArgPro Val Ser Ala Ala Ile Arg Glu Phe Phe Gly Thr 410 415 420 tcc cag ctgtct cag ttc atg gac cag aac aac tcc ctg tct ggt ttg 2012 Ser Gln Leu SerGln Phe Met Asp Gln Asn Asn Ser Leu Ser Gly Leu 425 430 435 act tac aagcgt cgt ctg tcg gct ctg ggc ccg ggt ggt ctg tcc cgt 2060 Thr Tyr Lys ArgArg Leu Ser Ala Leu Gly Pro Gly Gly Leu Ser Arg 440 445 450 gag cgc gccggc atc gag gtt cga gac gtt cac cca tct cac tac ggc 2108 Glu Arg Ala GlyIle Glu Val Arg Asp Val His Pro Ser His Tyr Gly 455 460 465 cgt atg tgccca att gag act ccg gaa ggt cca aac att ggc ctg atc 2156 Arg Met Cys ProIle Glu Thr Pro Glu Gly Pro Asn Ile Gly Leu Ile 470 475 480 485 ggt tccttg gct tcc tat gct cga gtg aac cca ttc ggt ttc att gag 2204 Gly Ser LeuAla Ser Tyr Ala Arg Val Asn Pro Phe Gly Phe Ile Glu 490 495 500 acc ccatac cgt cgc atc atc gac ggc aag ctg acc gac cag att gac 2252 Thr Pro TyrArg Arg Ile Ile Asp Gly Lys Leu Thr Asp Gln Ile Asp 505 510 515 tac cttacc gct gat gag gaa gac cgc ttc gtt gtt gcg cag gca aac 2300 Tyr Leu ThrAla Asp Glu Glu Asp Arg Phe Val Val Ala Gln Ala Asn 520 525 530 acg cactac gac gaa gag ggc aac atc acc gat gag acc gtc act gtt 2348 Thr His TyrAsp Glu Glu Gly Asn Ile Thr Asp Glu Thr Val Thr Val 535 540 545 cgt ctgaag gac ggc gac atc gcc atg gtt ggc cgc aac gcg gtt gat 2396 Arg Leu LysAsp Gly Asp Ile Ala Met Val Gly Arg Asn Ala Val Asp 550 555 560 565 tacatg gac gtt tcc cct cgt cag atg gtt tct gtt ggt acc gcg atg 2444 Tyr MetAsp Val Ser Pro Arg Gln Met Val Ser Val Gly Thr Ala Met 570 575 580 attcca ttc ctg gag cac gac gat gct aac cgt gca ctg atg ggc gcg 2492 Ile ProPhe Leu Glu His Asp Asp Ala Asn Arg Ala Leu Met Gly Ala 585 590 595 aacatg cag aag cag gct gtg cca ctg att cgt gcc gag gct cct ttc 2540 Asn MetGln Lys Gln Ala Val Pro Leu Ile Arg Ala Glu Ala Pro Phe 600 605 610 gtgggc acc ggt atg gag cag cgc gca gca tac gac gcc ggc gac ctg 2588 Val GlyThr Gly Met Glu Gln Arg Ala Ala Tyr Asp Ala Gly Asp Leu 615 620 625 gttatt acc cca gtc gca ggt gtg gtg gaa aac gtt tca gct gac ttc 2636 Val IleThr Pro Val Ala Gly Val Val Glu Asn Val Ser Ala Asp Phe 630 635 640 645atc acc atc atg gct gat gac ggc aag cgc gaa acc tac ctg ctg cgt 2684 IleThr Ile Met Ala Asp Asp Gly Lys Arg Glu Thr Tyr Leu Leu Arg 650 655 660aag ttc cag cgc acc aac cag ggc acc agc tac aac cag aag cct ttg 2732 LysPhe Gln Arg Thr Asn Gln Gly Thr Ser Tyr Asn Gln Lys Pro Leu 665 670 675gtt aac ttg ggc gag cgc gtt gaa gct ggc cag gtt att gct gat ggt 2780 ValAsn Leu Gly Glu Arg Val Glu Ala Gly Gln Val Ile Ala Asp Gly 680 685 690cca ggt acc ttc aat ggt gaa atg tcc ctt ggc cgt aac ctt ctg gtt 2828 ProGly Thr Phe Asn Gly Glu Met Ser Leu Gly Arg Asn Leu Leu Val 695 700 705gcg ttc atg cct tgg gaa ggc cac aac tac gag gat gcg atc atc ctc 2876 AlaPhe Met Pro Trp Glu Gly His Asn Tyr Glu Asp Ala Ile Ile Leu 710 715 720725 aac cag aac atc gtt gag cag gac atc ttg acc tcg atc cac atc gag 2924Asn Gln Asn Ile Val Glu Gln Asp Ile Leu Thr Ser Ile His Ile Glu 730 735740 gag cac gag atc gat gcc cgc gac act aag ctt ggc gcc gaa gaa atc 2972Glu His Glu Ile Asp Ala Arg Asp Thr Lys Leu Gly Ala Glu Glu Ile 745 750755 acc cgc gac atc cct aat gtg tct gaa gaa gtc ctc aag gac ctc gac 3020Thr Arg Asp Ile Pro Asn Val Ser Glu Glu Val Leu Lys Asp Leu Asp 760 765770 gac cgc ggt att gtc cgc atc ggt gct gat gtt cgt gac ggc gac atc 3068Asp Arg Gly Ile Val Arg Ile Gly Ala Asp Val Arg Asp Gly Asp Ile 775 780785 ctg gtc ggt aag gtc acc cct aag ggc gag acc gag ctc acc ccg gaa 3116Leu Val Gly Lys Val Thr Pro Lys Gly Glu Thr Glu Leu Thr Pro Glu 790 795800 805 gag cgc ttg ctg cgc gca atc ttc ggt gag aag gcc cgc gaa gtt cgc3164 Glu Arg Leu Leu Arg Ala Ile Phe Gly Glu Lys Ala Arg Glu Val Arg 810815 820 gat acc tcc atg aag gtg cct cac ggt gag acc ggc aag gtc atc ggc3212 Asp Thr Ser Met Lys Val Pro His Gly Glu Thr Gly Lys Val Ile Gly 825830 835 gtg cgt cac ttc tcc cgc gag gac gac gac gat ctg gct cct ggc gtc3260 Val Arg His Phe Ser Arg Glu Asp Asp Asp Asp Leu Ala Pro Gly Val 840845 850 aac gag atg atc cgt atc tac gtt gct cag aag cgt aag atc cag gac3308 Asn Glu Met Ile Arg Ile Tyr Val Ala Gln Lys Arg Lys Ile Gln Asp 855860 865 ggc gat aag ctc gct ggc cgc cac ggt aac aag ggt gtt gtc ggt aaa3356 Gly Asp Lys Leu Ala Gly Arg His Gly Asn Lys Gly Val Val Gly Lys 870875 880 885 att ttg cct cag gaa gat atg cca ttc ctt cca gac ggc act cctgtt 3404 Ile Leu Pro Gln Glu Asp Met Pro Phe Leu Pro Asp Gly Thr Pro Val890 895 900 gac atc atc ttg aac acc cac ggt gtt cca cgt cgt atg aac attggt 3452 Asp Ile Ile Leu Asn Thr His Gly Val Pro Arg Arg Met Asn Ile Gly905 910 915 cag gtt ctt gag acc cac ctt ggc tgg ctg gca tct gct ggt tggtcc 3500 Gln Val Leu Glu Thr His Leu Gly Trp Leu Ala Ser Ala Gly Trp Ser920 925 930 gtg gat cct gaa gat cct gag aac gct gag ctc gtc aag act ctgcct 3548 Val Asp Pro Glu Asp Pro Glu Asn Ala Glu Leu Val Lys Thr Leu Pro935 940 945 gca gac ctc ctc gag gtt cct gct ggt tcc ttg act gca act cctgtg 3596 Ala Asp Leu Leu Glu Val Pro Ala Gly Ser Leu Thr Ala Thr Pro Val950 955 960 965 ttc gac ggt gcg tca aac gaa gag ctc gca ggc ctg ctc gctaat tca 3644 Phe Asp Gly Ala Ser Asn Glu Glu Leu Ala Gly Leu Leu Ala AsnSer 970 975 980 cgt cca aac cgc gac ggc gac gtc atg gtt aac gcg gat ggtaaa gca 3692 Arg Pro Asn Arg Asp Gly Asp Val Met Val Asn Ala Asp Gly LysAla 985 990 995 acg ctt atc gac ggt cgc tcc ggt gag cct tac ccg tac ccggtt 3737 Thr Leu Ile Asp Gly Arg Ser Gly Glu Pro Tyr Pro Tyr Pro Val1000 1005 1010 tcc atc ggc tac atg tac atg ctg aag ctg cac cac ctc gttgac 3782 Ser Ile Gly Tyr Met Tyr Met Leu Lys Leu His His Leu Val Asp1015 1020 1025 gag aag atc cac gca cgt tcc act ggt cct tac tcc atg attacc 3827 Glu Lys Ile His Ala Arg Ser Thr Gly Pro Tyr Ser Met Ile Thr1030 1035 1040 cag cag cca ctg ggt ggt aaa gca cag ttc ggt gga cag cgtttc 3872 Gln Gln Pro Leu Gly Gly Lys Ala Gln Phe Gly Gly Gln Arg Phe1045 1050 1055 ggc gaa atg gag gtg tgg gca atg cag gca tac ggc gct gcctac 3917 Gly Glu Met Glu Val Trp Ala Met Gln Ala Tyr Gly Ala Ala Tyr1060 1065 1070 aca ctt cag gag ctg ctg acc atc aag tct gat gac gtg gttggc 3962 Thr Leu Gln Glu Leu Leu Thr Ile Lys Ser Asp Asp Val Val Gly1075 1080 1085 cgt gtc aag gtc tac gaa gca att gtg aag ggc gag aac atcccg 4007 Arg Val Lys Val Tyr Glu Ala Ile Val Lys Gly Glu Asn Ile Pro1090 1095 1100 gat cca ggt att cct gag tcc ttc aag gtt ctc ctc aag gagctc 4052 Asp Pro Gly Ile Pro Glu Ser Phe Lys Val Leu Leu Lys Glu Leu1105 1110 1115 cag tcc ttg tgc ctg aac gtg gag gtt ctc tcc gca gac ggcact 4097 Gln Ser Leu Cys Leu Asn Val Glu Val Leu Ser Ala Asp Gly Thr1120 1125 1130 cca atg gag ctc gcg ggt gac gac gac gac ttc gat cag gcaggc 4142 Pro Met Glu Leu Ala Gly Asp Asp Asp Asp Phe Asp Gln Ala Gly1135 1140 1145 gcc tca ctt ggc atc aac ctg tcc cgt gac gag cgt tcc gacgcc 4187 Ala Ser Leu Gly Ile Asn Leu Ser Arg Asp Glu Arg Ser Asp Ala1150 1155 1160 gac acc gca tagcagatca gaaaacaacc gctagaaatc aagccataca4236 Asp Thr Ala 1165 tcccccggac attgaagaga tgttctgggg ggaaagggagttttacgtgc tcgacgtaaa 4296 cgtcttcgat gagctccgca tcggcctggc caccgccgacgacatccgcc gttggtccaa 4356 gggtgaggtc aagaagccgg agaccatcaa ctaccgaaccctcaagcctg agaaggacgg 4416 tctgttctgc gagcgtatct tcggtccaac tcgcgactgggagtgcgcct gcggtaagta 4476 caagcgtgtc cgctacaagg gcatcatctg tgaacgctgtggcgttgagg tcaccaagtc 4536 caaggtgcgc cgtgagcgca tgggacacat tgagctcgctgcaccagtaa cccacatttg 4596 gtacttcaag ggcgttccat cacgcctcgg ctaccttttggaccttgctc caaaggacct 4656 ggacctcatc atctacttcg gtgcgaacat catcaccagcgtggacgaag aggctcgcca 4716 cagcgaccag accactcttg aggcagaaat gcttctggagaagaaggacg ttgaggcaga 4776 cgcagagtct gacattgctg agcgtgctga aaagctcgaagaggatcttg ctgaacttga 4836 ggcagctggc gctaaggccg acgctcgccg caaggttcaggctgctgccg ataaggaaat 4896 gcagcacatc cgtgagcgtg cacagcgcga aatcgatcgtctcgatgagg tctggcagac 4956 cttcatcaag cttgctccaa agcagatgat ccgcgatgagaagctctacg atgaactgat 5016 cgaccgctac gaggattact tcaccggtgg tatgggtgcagagtccattg aggctttgat 5076 ccagaacttc gaccttgatg ctg 5099 6 1165 PRTCorynebacterium glutamicum 6 Val Leu Glu Gly Pro Ile Leu Ala Val Ser ArgGln Thr Lys Ser Val 1 5 10 15 Val Asp Ile Pro Gly Ala Pro Gln Arg TyrSer Phe Ala Lys Val Ser 20 25 30 Ala Pro Ile Glu Val Pro Gly Leu Leu AspLeu Gln Leu Asp Ser Tyr 35 40 45 Ser Trp Leu Ile Gly Thr Pro Glu Trp ArgAla Arg Gln Lys Glu Glu 50 55 60 Phe Gly Glu Gly Ala Arg Val Thr Ser GlyLeu Glu Asn Ile Leu Glu 65 70 75 80 Glu Leu Ser Pro Ile Gln Asp Tyr SerGly Asn Met Ser Leu Ser Leu 85 90 95 Ser Glu Pro Arg Phe Glu Asp Val LysAsn Thr Ile Asp Glu Ala Lys 100 105 110 Glu Lys Asp Ile Asn Tyr Ala AlaPro Leu Tyr Val Thr Ala Glu Phe 115 120 125 Val Asn Asn Thr Thr Gly GluIle Lys Ser Gln Thr Val Phe Ile Gly 130 135 140 Asp Phe Pro Met Met ThrAsp Lys Gly Thr Phe Ile Ile Asn Gly Thr 145 150 155 160 Glu Arg Val ValVal Ser Gln Leu Val Arg Ser Pro Gly Val Tyr Phe 165 170 175 Asp Gln ThrIle Asp Lys Ser Thr Glu Arg Pro Leu His Ala Val Lys 180 185 190 Val IlePro Ser Arg Gly Ala Trp Leu Glu Phe Asp Val Asp Lys Arg 195 200 205 AspSer Val Gly Val Arg Ile Asp Arg Lys Arg Arg Gln Pro Val Thr 210 215 220Val Leu Leu Lys Ala Leu Gly Trp Thr Thr Glu Gln Ile Thr Glu Arg 225 230235 240 Phe Gly Phe Ser Glu Ile Met Met Ser Thr Leu Glu Ser Asp Gly Val245 250 255 Ala Asn Thr Asp Glu Ala Leu Leu Glu Ile Tyr Arg Lys Gln ArgPro 260 265 270 Gly Glu Gln Pro Thr Arg Asp Leu Ala Gln Ser Leu Leu AspAsn Ser 275 280 285 Phe Phe Arg Ala Lys Arg Tyr Asp Leu Ala Arg Val GlyArg Tyr Lys 290 295 300 Ile Asn Arg Lys Leu Gly Leu Gly Gly Asp His AspGly Leu Met Thr 305 310 315 320 Leu Thr Glu Glu Asp Ile Ala Thr Thr IleGlu Tyr Leu Val Arg Leu 325 330 335 His Ala Gly Glu Arg Val Met Thr SerPro Asn Gly Glu Glu Ile Pro 340 345 350 Val Glu Thr Asp Asp Ile Asp HisPhe Gly Asn Arg Arg Leu Arg Thr 355 360 365 Val Gly Glu Leu Ile Gln AsnGln Val Arg Val Gly Leu Ser Arg Met 370 375 380 Glu Arg Val Val Arg GluArg Met Thr Thr Gln Asp Ala Glu Ser Ile 385 390 395 400 Thr Pro Thr SerLeu Ile Asn Val Arg Pro Val Ser Ala Ala Ile Arg 405 410 415 Glu Phe PheGly Thr Ser Gln Leu Ser Gln Phe Met Asp Gln Asn Asn 420 425 430 Ser LeuSer Gly Leu Thr Tyr Lys Arg Arg Leu Ser Ala Leu Gly Pro 435 440 445 GlyGly Leu Ser Arg Glu Arg Ala Gly Ile Glu Val Arg Asp Val His 450 455 460Pro Ser His Tyr Gly Arg Met Cys Pro Ile Glu Thr Pro Glu Gly Pro 465 470475 480 Asn Ile Gly Leu Ile Gly Ser Leu Ala Ser Tyr Ala Arg Val Asn Pro485 490 495 Phe Gly Phe Ile Glu Thr Pro Tyr Arg Arg Ile Ile Asp Gly LysLeu 500 505 510 Thr Asp Gln Ile Asp Tyr Leu Thr Ala Asp Glu Glu Asp ArgPhe Val 515 520 525 Val Ala Gln Ala Asn Thr His Tyr Asp Glu Glu Gly AsnIle Thr Asp 530 535 540 Glu Thr Val Thr Val Arg Leu Lys Asp Gly Asp IleAla Met Val Gly 545 550 555 560 Arg Asn Ala Val Asp Tyr Met Asp Val SerPro Arg Gln Met Val Ser 565 570 575 Val Gly Thr Ala Met Ile Pro Phe LeuGlu His Asp Asp Ala Asn Arg 580 585 590 Ala Leu Met Gly Ala Asn Met GlnLys Gln Ala Val Pro Leu Ile Arg 595 600 605 Ala Glu Ala Pro Phe Val GlyThr Gly Met Glu Gln Arg Ala Ala Tyr 610 615 620 Asp Ala Gly Asp Leu ValIle Thr Pro Val Ala Gly Val Val Glu Asn 625 630 635 640 Val Ser Ala AspPhe Ile Thr Ile Met Ala Asp Asp Gly Lys Arg Glu 645 650 655 Thr Tyr LeuLeu Arg Lys Phe Gln Arg Thr Asn Gln Gly Thr Ser Tyr 660 665 670 Asn GlnLys Pro Leu Val Asn Leu Gly Glu Arg Val Glu Ala Gly Gln 675 680 685 ValIle Ala Asp Gly Pro Gly Thr Phe Asn Gly Glu Met Ser Leu Gly 690 695 700Arg Asn Leu Leu Val Ala Phe Met Pro Trp Glu Gly His Asn Tyr Glu 705 710715 720 Asp Ala Ile Ile Leu Asn Gln Asn Ile Val Glu Gln Asp Ile Leu Thr725 730 735 Ser Ile His Ile Glu Glu His Glu Ile Asp Ala Arg Asp Thr LysLeu 740 745 750 Gly Ala Glu Glu Ile Thr Arg Asp Ile Pro Asn Val Ser GluGlu Val 755 760 765 Leu Lys Asp Leu Asp Asp Arg Gly Ile Val Arg Ile GlyAla Asp Val 770 775 780 Arg Asp Gly Asp Ile Leu Val Gly Lys Val Thr ProLys Gly Glu Thr 785 790 795 800 Glu Leu Thr Pro Glu Glu Arg Leu Leu ArgAla Ile Phe Gly Glu Lys 805 810 815 Ala Arg Glu Val Arg Asp Thr Ser MetLys Val Pro His Gly Glu Thr 820 825 830 Gly Lys Val Ile Gly Val Arg HisPhe Ser Arg Glu Asp Asp Asp Asp 835 840 845 Leu Ala Pro Gly Val Asn GluMet Ile Arg Ile Tyr Val Ala Gln Lys 850 855 860 Arg Lys Ile Gln Asp GlyAsp Lys Leu Ala Gly Arg His Gly Asn Lys 865 870 875 880 Gly Val Val GlyLys Ile Leu Pro Gln Glu Asp Met Pro Phe Leu Pro 885 890 895 Asp Gly ThrPro Val Asp Ile Ile Leu Asn Thr His Gly Val Pro Arg 900 905 910 Arg MetAsn Ile Gly Gln Val Leu Glu Thr His Leu Gly Trp Leu Ala 915 920 925 SerAla Gly Trp Ser Val Asp Pro Glu Asp Pro Glu Asn Ala Glu Leu 930 935 940Val Lys Thr Leu Pro Ala Asp Leu Leu Glu Val Pro Ala Gly Ser Leu 945 950955 960 Thr Ala Thr Pro Val Phe Asp Gly Ala Ser Asn Glu Glu Leu Ala Gly965 970 975 Leu Leu Ala Asn Ser Arg Pro Asn Arg Asp Gly Asp Val Met ValAsn 980 985 990 Ala Asp Gly Lys Ala Thr Leu Ile Asp Gly Arg Ser Gly GluPro Tyr 995 1000 1005 Pro Tyr Pro Val Ser Ile Gly Tyr Met Tyr Met LeuLys Leu His 1010 1015 1020 His Leu Val Asp Glu Lys Ile His Ala Arg SerThr Gly Pro Tyr 1025 1030 1035 Ser Met Ile Thr Gln Gln Pro Leu Gly GlyLys Ala Gln Phe Gly 1040 1045 1050 Gly Gln Arg Phe Gly Glu Met Glu ValTrp Ala Met Gln Ala Tyr 1055 1060 1065 Gly Ala Ala Tyr Thr Leu Gln GluLeu Leu Thr Ile Lys Ser Asp 1070 1075 1080 Asp Val Val Gly Arg Val LysVal Tyr Glu Ala Ile Val Lys Gly 1085 1090 1095 Glu Asn Ile Pro Asp ProGly Ile Pro Glu Ser Phe Lys Val Leu 1100 1105 1110 Leu Lys Glu Leu GlnSer Leu Cys Leu Asn Val Glu Val Leu Ser 1115 1120 1125 Ala Asp Gly ThrPro Met Glu Leu Ala Gly Asp Asp Asp Asp Phe 1130 1135 1140 Asp Gln AlaGly Ala Ser Leu Gly Ile Asn Leu Ser Arg Asp Glu 1145 1150 1155 Arg SerAsp Ala Asp Thr Ala 1160 1165 7 1775 DNA Corynebacterium glutamicum CDS(500)..(880) 7 cagctctaca agagtgtcta agtggcgggc attccatgct ttggaggagcgatcttcaaa 60 ttcctccaaa gtgagttgac ctcgggaaac agctgcagaa agttcatccacgacttggtt 120 tcggttaagg tcagtggcga gcttctttgc tggttcgttt ccttgaggaacagtcatggg 180 aaccattcta acaagggatt tggtgttttc tgcggctagc tgataatgtgaacggctgag 240 tcccactctt gtagttggga attgacggca cctcgcactc aagcgcggtatcgcccctgg 300 ttttccggga cgcggtggcg catgtttgca tttgatgagg ttgtccgtgacatgtttggt 360 cgggccccaa aaagagcccc cttttttgcg tgtctggaca ctttttcaaatccttcgcca 420 tcgacaagct cagccttcgt gttcgtcccc cgggcgtcac gtcagcagttaaagaacaac 480 tccgaaataa ggatggttc atg cca act att cag cag ctg gtc cgtaag ggc 532 Met Pro Thr Ile Gln Gln Leu Val Arg Lys Gly 1 5 10 cgc cacgat aag tcc gcc aag gtg gct acc gcg gca ctg aag ggt tcc 580 Arg His AspLys Ser Ala Lys Val Ala Thr Ala Ala Leu Lys Gly Ser 15 20 25 cct cag cgtcgt ggc gta tgc acc cgt gtg tac acc acc acc cct aag 628 Pro Gln Arg ArgGly Val Cys Thr Arg Val Tyr Thr Thr Thr Pro Lys 30 35 40 aag cct aac tctgct ctt cgt aag gtc gct cgt gtg cgc ctt acc tcc 676 Lys Pro Asn Ser AlaLeu Arg Lys Val Ala Arg Val Arg Leu Thr Ser 45 50 55 ggc atc gag gtt tccgct tac atc cct ggt gag ggc cac aac ctg cag 724 Gly Ile Glu Val Ser AlaTyr Ile Pro Gly Glu Gly His Asn Leu Gln 60 65 70 75 gag cac tcc atg gtgctc gtt cgc ggt ggt cgt gtt aag gac ctc cca 772 Glu His Ser Met Val LeuVal Arg Gly Gly Arg Val Lys Asp Leu Pro 80 85 90 ggt gtc cgt tac aag atcgtc cgt ggc gca ctg gat acc cag ggt gtt 820 Gly Val Arg Tyr Lys Ile ValArg Gly Ala Leu Asp Thr Gln Gly Val 95 100 105 aag gac cgc aag cag gctcgt tcc ccg cta cgg cgc gaa gag ggg ata 868 Lys Asp Arg Lys Gln Ala ArgSer Pro Leu Arg Arg Glu Glu Gly Ile 110 115 120 Ile Lys Asn Ala 125gtatacaagt ccgagctcgt tacccagctc gtaaacaaga tcctcatcgg tggcaagaag 980tccaccgcag agcgcatcgt ctacggtgca ctcgagatct gccgtgagaa gaccggcacc 1040gatccagtag gaaccctcga gaaggctctc ggcaacgtgc gtccagacct cgaagttcgt 1100tcccgccgtg ttggtggcgc tacctaccag gtgccagtgg atgttcgccc agagcgcgca 1160aacaccctcg cactgcgttg gttggtaacc ttcacccgtc agcgtcgtga gaacaccatg 1220atcgagcgtc ttgcaaacga acttctggat gcagccaacg gccttggcgc ttccgtgaag 1280cgtcgcgaag acacccacaa gatggcagag gccaaccgcg ccttcgctca ctaccgctgg 1340tagtactgcc aagacatgaa agcccaatca cctttaagat caacgcctgc cggcgccctt 1400cacatttgaa taagctggca gcctgcgttt cttcaaggcg actgggcttt tagtctcatt 1460aatgcagttc accgctgtaa gatagctaaa tagaaacact gtttcggcag tgtgttacta 1520aaaaatccat gtcacttgcc tcgagcgtgc tgcttgaatc gcaagttagt ggcaaaatgt 1580aacaagagaa ttatccgtag gtgacaaact ttttaatact tgggtatctg tcatggatac 1640cccggtaata aataagtgaa ttaccgtaac caacaagttg gggtaccact gtggcacaag 1700aagtgcttaa ggatctaaac aaggtccgca acatcggcat catggcgcac atcgatgctg 1760gtaagaccac gacca 1775 8 127 PRT Corynebacterium glutamicum 8 Met Pro ThrIle Gln Gln Leu Val Arg Lys Gly Arg His Asp Lys Ser 1 5 10 15 Ala LysVal Ala Thr Ala Ala Leu Lys Gly Ser Pro Gln Arg Arg Gly 20 25 30 Val CysThr Arg Val Tyr Thr Thr Thr Pro Lys Lys Pro Asn Ser Ala 35 40 45 Leu ArgLys Val Ala Arg Val Arg Leu Thr Ser Gly Ile Glu Val Ser 50 55 60 Ala TyrIle Pro Gly Glu Gly His Asn Leu Gln Glu His Ser Met Val 65 70 75 80 LeuVal Arg Gly Gly Arg Val Lys Asp Leu Pro Gly Val Arg Tyr Lys 85 90 95 IleVal Arg Gly Ala Leu Asp Thr Gln Gly Val Lys Asp Arg Lys Gln 100 105 110Ala Arg Ser Pro Leu Arg Arg Glu Glu Gly Ile Ile Lys Asn Ala 115 120 1259 24 DNA ARTIFICIAL SEQUENCE SYNTHETIC DNA 9 acaatgtgac tcgtgatttt tggg24 10 20 DNA ARTIFICIAL SEQUENCE SYNTHETIC DNA 10 ggaaacgtcc atgtaatcaa20 11 20 DNA ARTIFICIAL SEQUENCE SYNTHETIC DNA 11 aacacgcact acgacgaaga20 12 20 DNA ARTIFICIAL SEQUENCE SYNTHETIC DNA 12 cagcatcaag gtcgaagttc20

1. An isolated polynucleotide which encodes a protein comprising theamino acid sequence of SEQ ID NO:2.
 2. The isolated polynucleotide ofclaim 1, wherein said protein has the activity of the β-subunit of RNApolymerase B.
 3. An isolated polynucleotide which comprises SEQ ID NO:1.4. An isolated polynucleotide which is complimentary to thepolynucleotide of claim
 3. 5. An isolated polynucleotide which is atleast 70% identical to the polynucleotide of claim
 3. 6. An isolatedpolynucleotide which is at least 80% identical to the polynucleotide ofclaim
 3. 7. An isolated polynucleotide which is at least 90% identicalto the polynucleotide of claim
 3. 8. An isolated polynucleotide whichhybridizes under stringent conditions to the polynucleotide of claim 3;wherein said stringent conditions comprise washing in 5×SSC at atemperature from 50 to 68° C.
 9. The isolated polynucleotide of claim 3,which encodes a protein having the activity of the β-subunit of RNApolymerase B.
 10. An isolated polynucleotide which comprises at least 15consecutive nucleotides of the polynucleotide of claim
 3. 11. Anisolated polypeptide which comprises the amino acid sequence of SEQ IDNO:2.
 12. An isolated polypeptide which comprises the amino acidsequence of SEQ ID NO:4.
 13. An isolated polypeptide which comprises theamino acid sequence of SEQ ID NO:6.
 14. An isolated polynucleotide whichencodes a protein comprising the amino acid sequence of SEQ ID NO:4. 15.An isolated polynucleotide which comprises SEQ ID NO:3.
 16. An isolatedpolynucleotide which encodes a protein comprising the amino acidsequence of SEQ ID NO:6.
 17. An isolated polynucleotide which comprisesSEQ ID NO:5.
 18. A vector comprising the isolated polynucleotide ofclaim
 1. 19. A vector comprising the isolated polynucleotide of claim 3.20. A vector comprising the isolated polynucleotide of claim
 14. 21. Avector comprising the isolated polynucleotide of claim
 15. 22. A vectorcomprising the isolated polynucleotide of claim
 16. 23. A vectorcomprising the isolated polynucleotide of claim
 17. 24. A host cellcomprising the isolated polynucleotide of claim
 1. 25. A host cellcomprising the isolated polynucleotide of claim
 3. 26. A host cellcomprising the isolated polyncleotide of claim
 14. 27. A host cellcomprising the isolated polyncleotide of claim
 15. 28. A host cellcomprising the isolated polyncleotide of claim
 16. 29. A host cellcomprising the isolated polyncleotide of claim
 17. 30. The host cell ofclaim 24, which is a Coryneform bacterium.
 31. The host cell of claim25, which is a Coryneform bacterium.
 32. The host cell of claim 26,which is a Coryneform bacterium.
 33. The host cell of claim 27, which isa Coryneform bacterium.
 34. The host cell of claim 28, which is aCoryneform bacterium.
 35. The host cell of claim 29, which is aCoryneform bacterium.
 36. The host cell of claim 24, wherein said hostcell is selected from the group consisting of Coryneform glutamicum,Corynebacterium acetoglutamicum, Corynebacterium acetoacidophilum,Corynebacterium thermoaminogenes, Corynebacterium melassecola,Brevibacterium flavum, Brevibacterium lactofermentum, and Brevibacteriumdivaricatum.
 37. The host cell of claim 24, wherein said host cell isselected from the group consisting of Corynebacterium glutamicum FERM1709, Brevibacterium flavum FERM-P 1708, Brevibacterium lactofermentumFERM-P1712, Corynebacterium glutamicum FERM-P6463, Corynebacteriumglutamicum FERM-P6464, Corynebacterium glutamicum DM58-1,Corynebacterium glutamicum DG 52-5, Corynebacterium glutamicum DSM 5714and Corynebacterium glutamicum DSM-12866.
 38. The host cell of claim 25,wherein said host cell is selected from the group consisting ofCoryneform glutamicum, Corynebacterium acetoglutamicum, Corynebacteriumacetoacidophilum, Corynebacterium thermoaminogenes, Corynebacteriummelassecola, Brevibacterium flavum, Brevibacterium lactofermentum, andBrevibacterium divaricatum.
 39. The host cell of claim 25, wherein saidhost cell is selected from the group consisting of Corynebacteriumglutamicum FERM 1709, Brevibacterium flavum FERM-P 1708,Brevibacterium.lactofermentum FERM-P1712, Corynebacterium glutamicumFERM-P6463, Corynebacterium glutamicum FERM-P6464, Corynebacteriumglutamicum DM58-1, Corynebacterium glutamicum DG 52-5, Corynebacteriumglutamicum DSM 5714 and Corynebacterium glutamicum DSM-12866.
 40. ACoryneform bacterium which comprises an enhanced rpoB gene.
 41. TheCoryneform bacterium of claim 40, wherein said rpoB gene comprises thepolynucleotide sequence of SEQ ID NO:1.
 42. The Coryneform bacterium ofclaim 40, wherein said enhanced rpoB gene comprises the polynucleotidesequence of SEQ ID NO:3.
 43. The Coryneform bacterium of claim 40,wherein said enhanced rpoB gene comprises the polynucleotide sequence ofSEQ ID NO:5.
 44. Coryneform glutamicum DSM
 13993. 45. Coryneformglutamicum DSM
 13994. 46. A process for producing L-amino acidscomprising culturing the host cell of claim 24 in a medium suitable forthe expression of the polynucleotide; and collecting the L-amino acid.47. The process of claim 46, wherein said L-amino acid is L-lysine orL-glutamate.
 48. The process of claim 46, wherein said L-amino acid isL-lysine and the host cell further comprises at least one gene whoseexpression is enhanced, wherein said gene is selected from the groupconsisting of dapA, gap, tpi, pgk, zwf, pyc, mqo, lys C, lys E, zwa1 andrpsl.
 49. The process of claim 46, wherein the host cell furthercomprises at least one gene whose expression is attenuated, wherein saidgene is selected from the group consisting of pck gene, pgi gene, poxB,and zwa2.
 50. A process for producing L-amino acids comprising culturingthe host cell of claim 25 in a medium suitable for the expression of thepolynucleotide; and collecting the L-amino acid.
 51. The process ofclaim 50, wherein said L-amino acid is L-lysine or L-glutamate.
 52. Theprocess of claim 50, wherein wherein said L-amino acid is L-lysine andthe host cell further comprises at least one gene whose expression isenhanced, wherein said gene is selected from the group consisting ofdapA, gap, tpi, pgk, zwf, pyc, mqo, lys C, lys E, zwa1 and rpsL.
 53. Theprocess of claim 50, wherein the host cell further comprises at leastone gene whose expression is attenuated, wherein said gene is selectedfrom the group consisting of pck gene, pgi gene, poxB, and zwa2.
 54. Aprocess for producing L-amino acids comprising culturing the host cellof claim 26 in a medium suitable for the expression of thepolynucleotide; and collecting the L-amino acid.
 55. The process ofclaim 54, wherein said L-amino acid is L-lysine or L-glutamate.
 56. Theprocess of claim 54, wherein wherein said L-amino acid is L-lysine andthe host cell further comprises at least one gene whose expression isenhanced, wherein said gene is selected from the group consisting ofdapA, gap, tpi, pgk, zwf, pyc, mqo, lys C, lys E, zwal and rpsL.
 57. Theprocess of claim 54, wherein the host cell further comprises at leastone gene whose expression is attenuated, wherein said gene is selectedfrom the group consisting of pck gene, pgi gene, poxB, and zwa2.
 58. Aprocess for producing L-amino acids comprising culturing the host cellof claim 26 in a medium suitable for the expression of thepolynucleotide; and collecting the L-amino acid.
 59. The process ofclaim 58, wherein said L-amino acid is L-lysine or L-glutamate.
 60. Theprocess of claim 58, wherein wherein said L-amino acid is L-lysine andthe host cell further comprises at least one gene whose expression isenhanced, wherein said gene is selected from the group consisting ofdapA, gap, tpi, pgk, zwf, pyc, mqo, lys C, lys E, zwa1 and rpsL.
 61. Theprocess of claim 58, wherein the host cell further comprises at leastone gene whose expression is attenuated, wherein said gene is selectedfrom the group consisting of pck gene, pgi gene, poxB, and zwa2.
 62. Aprocess for producing L-amino acids comprising culturing the host cellof claim 27 in a medium suitable for the expression of thepolynucleotide; and collecting the L-amino acid.
 63. The process ofclaim 62, wherein said L-amino acid is L-lysine or L-glutamate.
 64. Theprocess of claim 62, wherein wherein said L-amino acid is L-lysine andthe host cell further comprises at least one gene whose expression isenhanced, wherein said gene is selected from the group consisting ofdapA, gap, tpi, pgk, zwf, pyc, mqo, lys C, lys E, zwa1 and rpsL.
 65. Theprocess of claim 62, wherein the host cell further comprises at leastone gene whose expression is attenuated, wherein said gene is selectedfrom the group consisting of pck gene, pgi gene, poxB, and zwa2.
 66. Aprocess for producing L-amino acids comprising culturing the host cellof claim 28 in a medium suitable for the expression of thepolynucleotide; and collecting the L-amino acid.
 67. The process ofclaim 66, wherein said L-amino acid is L-lysine or L-glutamate.
 68. Theprocess of claim 66, wherein wherein said L-amino acid is L-lysine andthe host cell further comprises at least one gene whose expression isenhanced, wherein said gene is selected from the group consisting ofdapA, gap, tpi, pgk, zwf, pyc, mqo, lys C, lys E, zwa1 and rpsL.
 69. Theprocess of claim 66, wherein the host cell further comprises at leastone gene whose expression is attenuated, wherein said gene is selectedfrom the group consisting of pck gene, pgi gene, poxB, and zwa2.
 70. Aprocess for producing L-amino acids comprising culturing the host cellof claim 29 in a medium suitable for the expression of thepolynucleotide; and collecting the L-amino acid.
 71. The process ofclaim 70, wherein said L-amino acid is L-lysine or L-glutamate.
 72. Theprocess of claim 70, wherein wherein said L-amino acid is L-lysine andthe host cell further comprises at least one gene whose expression isenhanced, wherein said gene is selected from the group consisting ofdapA, gap, tpi, pgk, zwf, pyc, mqo, lys C, lys E, zwa1 and rpsL.
 73. Theprocess of claim 70, wherein the host cell further comprises at leastone gene whose expression is attenuated, wherein said gene is selectedfrom the group consisting of pck gene, pgi gene, poxB, and zwa2.
 74. Aprocess for screening for polynucleotides which encode a protein havingthe activity of the β-subunit of RNA polymerase B comprising hybridizingthe isolated polynucleotide of claim 1 to the polynucleotide to bescreened; expressing the polynucleotide to produce a protein; anddetecting the presence or absence of the activity of the β-subunit ofRNA polymerase B in said protein.
 75. A process for screening forpolynucleotides which encode a protein having the activity of theβ-subunit of RNA polymerase B comprising hybridizing the isolatedpolynucleotide of claim 3 to the polynucleotide to be screened;expressing the polynucleotide to produce a protein; and detecting thepresence or absence of the activity of the β-subunit of RNA polymerase Bin said protein.
 76. A process for screening for polynucleotides whichencode a protein having the activity of the β-subunit of RNA polymeraseB comprising hybridizing the isolated polynucleotide of claim 15 to thepolynucleotide to be screened; expressing the polynucleotide to producea protein; and detecting the presence or absence the activity of theβ-subunit of RNA polymerase B in said protein.
 77. A process forscreening for polynucleotides which encode a protein having the activityof the β-subunit of RNA polymerase B comprising hybridizing the isolatedpolynucleotide of claim 17 to the polynucleotide to be screened;expressing the polynucleotide to produce a protein; and detecting thepresence or absence the activity of the β-subunit of RNA polymerase B insaid protein.
 78. A method for detecting a nucleic acid with at least70% homology to nucleotide of claim 1, comprising contacting a nucleicacid sample with a probe or primer comprising at least 15 consecutivenucleotides of the nucleotide sequence of claim 1, or at least 15consecutive nucleotides of the complement thereof.
 79. A method forproducing a nucleic acid with at least 70% homology to nucleotide ofclaim 1, comprising contacting a nucleic acid sample with a primercomprising at least 15 consecutive nucleotides of the nucleotidesequence of claim 1, or at least 15 consecutive nucleotides of thecomplement thereof.
 80. A method for detecting a nucleic acid with atleast 70% homology to nucleotide of claim 3, comprising contacting anucleic acid sample with a probe or primer comprising at least 15consecutive nucleotides of the nucleotide sequence of claim 3, or atleast 15 consecutive nucleotides of the complement thereof.
 81. A methodfor producing a nucleic acid with at least 70% homology to nucleotide ofclaim 3, comprising contacting a nucleic acid sample with a primercomprising at least 15 consecutive nucleotides of the nucleotidesequence of claim 3, or at least 15 consecutive nucleotides of thecomplement thereof.
 82. A method for making a β-subunit of RNApolymerase B, comprising: culturing the host cell of claim 23 for a timeand under conditions suitable for expression of the β-subunit of RNApolymerase B, and collecting the β-subunit of RNA polymerase B.
 83. Amethod for making a β-subunit of RNA polymerase B, comprising: culturingthe host cell of claim 24 for a time and under conditions suitable forexpression of the β-subunit of RNA polymerase B, and collecting theβ-subunit of RNA polymerase B.
 84. A method for making a β-subunit ofRNA polymerase B, comprising: culturing the host cell of claim 25 for atime and under conditions suitable for expression of the β-subunit ofRNA polymerase B, and collecting the β-subunit of RNA polymerase B. 85.A method for making a β-subunit of RNA polymerase B, comprising:culturing the host cell of claim 26 for a time and under conditionssuitable for expression of the β-subunit of RNA polymerase B, andcollecting the β-subunit of RNA polymerase B.
 86. A method for making aβ-subunit of RNA polymerase B, comprising: culturing the host cell ofclaim 27 for a time and under conditions suitable for expression of theβ-subunit of RNA polymerase B, and collecting the β-subunit of RNApolymerase B.
 87. A method for making a β-subunit of RNA polymerase B,comprising: culturing the host cell of claim 28 for a time and underconditions suitable for expression of the β-subunit of RNA polymerase B,and collecting the β-subunit of RNA polymerase B.